The Biotechnology Education Company ®
sed
Revi nd
a
ated
Upd
EDVO-Kit #
119
The DNA
Report Card™
Storage: See Page 3 for
specific storage instructions
Experiment Objective:
The purpose of the experiment is to rapidly extract "self"
DNA from cheek cells, to visualize DNA, and to store
DNA on individual DNA Report Cards™.
EDVOTEK, Inc. • 1-800-EDVOTEK • www.edvotek.com
EVT 007176K
DNA Report Card™
EDVO-Kit #
119
xxx
Table of Contents
Page
Experiment Components
3
Experiment Requirements
3
Background Information
5
Experiment Procedures
Experiment Overview
Experiment Overview and General Instructions
Cheek Cell DNA Isolation
Preparing the DNA for Storage on the DNA Report Card
Preparation of the DNA Report Card
Study Questions
Instructor's Guidelines
Pre-Lab Preparations
Inquiry-based Optional Extension Experiments
Expected Results
Study Questions and Answers
Material Safety Data Sheets
8
9
10
11
12
13
15
16
17
17
18
19
All components are intended for
educational research only. They
are not to be used for diagnostic
or drug purposes, nor administered to or consumed by humans
or animals.
THIS EXPERIMENT DOES NOT
CONTAIN HUMAN DNA. None
of the experiment components
are derived from human sources.
EDVOTEK, The Biotechnology Education Company, and InstaStain are registered
trademarks of EDVOTEK, Inc.. Ready-to-Load and UltraSpec-Agarose are trademarks
of EDVOTEK, Inc.
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EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
119
Experiment Components
This experiment is for 26 DNA isolations.
Storage
•
•
•
•
•
Room temperature
Room temperature
Room temperature
Room temperature
Room temperature
Lysis Buffer
NaCl Solution
Protease
Tris Buffer
Methylene Blue Plus™ solution
--------------------------------------------------------------------•
•
•
•
•
•
DNA Report Cards™
Clear tubes for DNA isolation
Microcentrifuge tubes with caps
Sterile cotton tipped applicators
Small transfer pipets
Calibrated transfer pipets
Room temperature
Requirements
•
•
•
•
Freezer/ice cold 95% Ethanol
or isopropyl alcohol (rubbing alcohol)
Water bath (50 °C)
Test tube racks
Ice and ice buckets
• Disposable laboratory gloves
EDVOTEK - The Biotechnology Education Company®
1-800-EDVOTEK • www.edvotek.com
FAX: (301) 340-0582 • email: [email protected]
EVT 007176K
DNA Report Card™
EDVO-Kit #
119
xxx
Online Ordering
now available
E
O
DV
-TE
C H S E RV I C E
Technical Service
Department
1-800-EDVOTEK
(1-800-338-6835)
ET
Mo
Visit our web site for information
about EDVOTEK’s complete line
of “hands-on” experiments for
biotechnology and biology education.
Mon - Fri
9:00 am to 6:00 pm ET
FAX: (301) 340-0582
Web: www.edvotek.com
email: [email protected]
m
6p
n - Fri 9 am
Please have the following
information ready:
• Experiment number and title
• Kit lot number on box or tube
• Literature version number
(in lower right corner)
• Approximate purchase date
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DNA Report Card™
EDVO-Kit #
119
An Introduction About DNA
5'
T
A
C
G
T
A
C
G
A
T
G
C
Figure 1
The DNA structure was determined by James Watson
and Francis Crick in 1953. They determined that DNA
was a double helix consisting of two strands with opposite polarity. The Watson and Crick model is often
described as the spiral DNA ladder.
Human cells contain a nucleus, which contains 46 chromosomes (23 pairs). Chromosomes contain DNA which
encodes all the genetic information that is inherited
from the two biological parents. DNA is made up of
building blocks known as nucleotides. Each nucleotide
is composed of three parts, a phosphate group, deoxyribose sugar, and one of the four nitrogenous bases,
Adenine, Guanine, Cytosine, or Thymine.
C
G
3'
3'
5'
The two strands of DNA that make up the backbone of
the DNA ladder is made up of the deoxyribose carbohydrate units linked together by phosphodiester bonds.
The carbohydrate backbone acts as a support for the
rungs of the ladder. The rungs are composed of the
nitrogenous bases. The first letters of these bases, A, G,
C, and T, designate the order of the bases within the
two DNA strands. The four bases are always hydrogen
bonded in pairs. When A occurs on one strand, T will occur on the opposite strand. Similarly, G and C are base
pairs on opposite DNA strands. The bases are held together by hydrogen bonds which are shown as dashed
lines in Figure 1. Unlike covalent bonds hydrogen bonds
are much weaker.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
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The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
Background Information
All living organisms are composed of cells. Organisms such as bacteria are single cells, while very complex organisms, such as humans, are
composed of billions of many different cells. In 1868, a biologist named
Friedrich Meischer carried out research which indicated that the nucleus
of cells contains a material which he called nucleic acid. It was not until
much later in the 1940’s that deoxyribonucleic acid (DNA), was recognized as the carrier of the genetic code.
DNA Report Card™
EDVO-Kit #
119
An Introduction About DNA
Background Information
The human genome consists of 2.9 billion base pairs. Of this total, only
about 5% code for protein. Intervening sequences and other noncoding
sequences make up the remainder. Some of the noncoding sequences or
other functionally unassigned sequences may possess undiscovered functions. Certain noncoding sequences appear to be self-replicating and
are repeated hundreds or thousands of times throughout the genome.
These repetitive sequences have been termed “selfish” or “parasitic” DNA,
as they often appear to possess no function except that of
their own reproduction. Repetitive elements account for more than 20
percent of the human genome. In addition to genomic DNA, mitochondria, which are cellular organelles, contain their own DNA (mitochondrial
DNA) which replicate independently from cell chromosomal DNA.
During the process of replication, DNA provides the required information
to copy (replicate) itself, which results in genetic information to be passed
on to the next generation of cells. DNA also provides the instructions for
making proteins for various cell functions. A large number of proteins and
enzymes including DNA polymerases are involved in the synthesis of DNA.
In addition to synthesis DNA polymerases have the ability to edit newly
synthesized DNA for possible errors in base pair matching and to participate in repairing DNA that gets damaged during the life of the cell. Such
damage to DNA can occur due to exposure to carcinogens and environmental factors such as X-rays and U.V. light from exposure to the sun.
When cells are chemically lysed (broken open), DNA from
chromosomes is released and can be isolated and purified. DNA extraction is frequently the first step for molecular
biology and biotechnology experiments. Extracted DNA is
soluble in water and thus will appear as a clear solution. By
contrast DNA is insoluble in salt solutions and alcohol, where
it will form white fibers.
Figure 2
Purification procedures for DNA usually include precipitation
with alcohol in the presence of salt. The DNA solution is carefully overlaid with alcohol. Since alcohols such as rubbing
alcohol (isopropyl alcohol) have lower density than water a
second layer above the DNA solution will be formed. A glass
rod or a stirrer is used to spool DNA at the interface of the
two liquid phases and to separate the DNA from the solution (see Figure 2). The DNA will appear as a viscous, clotted
mass, which can be collected on a stirrer or a glass rod. The
amount of DNA spooled will vary and is a consequence of
the intactness of the DNA sample.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
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DNA Report Card™
EDVO-Kit #
119
An Introduction About DNA
A specified amount of DNA is required for DNA fingerprinting. Small
amounts of body fluids or tissues until recently was not sufficient for DNA
extraction and analysis. This is no longer a limitation if genes to be analyzed are first amplified by the polymerase chain reaction (PCR) prior to
use. In 1984 Kary Mullis invented PCR and for his unique contribution he
was awarded a Nobel Prize in 1994. The widespread utility of PCR is based
on ease of use and the ability to amplify DNA making it possible to perform a variety of studies including DNA fingerprinting. DNA amplified by
the PCR reaction is analyzed by agarose gel electrophoresis or become
part of additional procedures used in DNA studies.
In this experiment individual “self” DNA will be extracted from cheek cells
and stored in the EDVOTEK DNA Report Card™. DNA from the DNA Report Card™ can be recovered and used for DNA fingerprinting and analysis of an individual's genetics.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
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The Experiment
Almost any tissue or body fluid (except urine) may be used as a source
of DNA. The most common sources of human DNA are samples from hair,
cheek cells, blood and saliva. Once extracted, DNA can be stored for
long periods of time. Various methods of storage include precipitation and
storage under alcohol at room temperature or under refrigeration. DNA
can also be stored on the DNA Report Card™. DNA can be recovered
from the DNA Report Card™, small blood spots and tissues, or even a
few cells. Such amounts of DNA can be obtained from individuals during medical procedures or evidence left behind in crime scenes such as
cells that are recovered from the fingernails of a victim. In recent times,
a few cells deposited by a person while licking and sealing an envelope
has been sufficient to obtain DNA and match the DNA fingerprint to the
person who left this evidence behind.
DNA Report Card™
EDVO-Kit #
119
The Experiment
Experiment Overview
1
2
Extract DNA
Twirl
applicator
in lysis
buffer
(repeat
once)
Swab cheek
cells vigorously
4
3
5
Cap tube
and Invert
to mix
Add 22drops
drops of
of Protease
Protease
6
incubate
10 min.
Add 2 drops
Add
4 drops
of
NaCl
of NaCl
56°C
7
Cap tube
and Invert
to mix
8
Incubate
4 min.
Room
Temp.
11
10
9
56°C
Add 1 ml ice
cold ETHANOL
Let it stream
down side of
tube.
Allow
95% ethanol
or tube
91%
to sit upright.
isopropyl/rubbing
Allow
tube to sit
upright.
11
12
EDVO-Kit
®
Look
Lookfor
for
DNA
DNAatat
the
the
interface.
interface.
Look for
stringy/
white
material.
Gently invert
tube once
This is your DNA!!
# 119
DNA Report
Card™
Name: ________________
Date: __________________
Control
(No DNA)
Unstained
DNA
Methylene
Blue
Stained
DNA
Transfer the
DNA to the
DNA Report
card and/or a
pendant
(before or after
staining with
Methylene
Blue).
Copyright © 2004, EDVOTEK, Inc.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
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The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
119
Experiment Overview and General Instructions
Experiment Objective:
The purpose of the experiment is to rapidly extract "self" DNA from cheek
cells, to visualize DNA, and to store DNA on individual DNA Report
Cards™.
1. Gloves and goggles should be worn routinely as good laboratory
practice.
2. Exercise extreme caution when working with equipment that is used in
conjunction with the heating and/or melting of reagents.
3. Do not mouth pipet reagents - use pipet pumps.
4. Exercise caution when using any electrical
equipment in the laboratory.
5. Always wash hands thoroughly with soap
and water after handling reagents or biological materials in the laboratory.
Laboratory notebook recordings:
Address and record the following in your laboratory notebook or on a
separate worksheet.
Before starting the Experiment:
•
•
Write a hypothesis that reflects the experiment.
Predict experimental outcomes.
During the Experiment:
• Record (draw) your observations, or photograph the results.
Following the Experiment:
• Formulate an explanation from the results.
• Determine what could be changed in the experiment if the experiment were repeated.
• Write a hypothesis that would reflect this change.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
The Experiment
LABORATORY SAFETY
10
DNA Report Card™
EDVO-Kit #
119
Cheek Cell DNA Isolation
1. Obtain a clean capped tube containing 600 µl of lysis
buffer. Write your initials on the side of the tube with a lab
marker.
The Experiment
Using one of the sterile cotton-tipped applicators, collect
cheek cells by vigorously swabbing inside the mouth
along the gum line and under the tongue.
2. Place the applicator in the tube containing the Lysis buffer and twirl to dislodge the cells. Press the tip to the sides
of the tube to squeeze out as much liquid as possible.
Repeat steps 1 and 3 using a fresh sterile applicator. Twirl the applicator in the same tube containing lysis buffer to add cells to the solution.
3. Using a small transfer pipet, add 2 drops of the Protease solution to the
tube.
4. Cap and gently invert the tube several times to mix. Empty and save
the pipet for future use.
5. Incubate the tube in a 56°C waterbath for 10 minutes.
(The protease is most active at 56°C.)
6. Using a new small transfer pipet, add 2 drops of NaCl solution to the
tube.
7. Cap the tube and gently flick the tube with your index finger several
times to mix. Empty and save the pipet for future use (step 16).
8. Incubate the tube at room temperature for 4 minutes.
9. Using a clean calibrated transfer pipet, add 1 ml of ice cold alcohol to
the test tube. Hold the test tube at a 45° angle and gently pipet the
cold alcohol allowing it to flow down the side of the test tube.
Place the tube in a test tube rack sitting upright. Allow the tube to sit
for 3-5 minutes.
10. Check if your DNA is visible at the interface of the alcohol/salt solution.
Some DNA may appear as stringy material, grayish-white in color.
11. Cap the tube and gently invert it to mix the DNA solution and alcohol phases. Look for any stringy white
or clear material. This is your DNA.
12. Proceed to preparing the DNA for storage on the
DNA Report Card.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
11
119
Preparing the DNA for Storage on the DNA Report Card
Transfer Unstained Precipitated DNA
14. Use the pipet from step 5 and remove 2 drops of the white precipitate
and transfer to a clean microtest tube.
15. Label the tube "Unstained DNA" and save this DNA for transfer to the
DNA Report Card™.
16. Transfer some unstained DNA to a separate tube, and without disturbing the tube, use the transfer pipet from step 7 to add 1 drop of
Methylene Blue Plus™ solution. Empty and save the pipet for the next
step (step 17). Observe your DNA in the tube.
Methylene blue is a DNA stain which will make the extracted DNA more
easily visible to the naked eye. Your DNA will appear as heavy blue gel-like
strands at the alcohol and aqueous interface. The solution will also have a
light blue color but will be lighter than your DNA.
17. Transfer 1 drop of the Methylene Blue stained DNA to a clean microtest tube.
18. Label the tube "Stained DNA" and save this DNA for transfer to the
DNA Report Card™.
STORAGE OF Your precipitated DNA
19. After removing the portions of your unstained and stained DNA for
transfer to the DNA Report Card™, recap the tube containing your
precipitated DNA.
20. Label the tube with your initials.
21. Place the DNA in the freezer and observe the DNA in the tube the
next day or at a later time.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
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The Experiment
Stain the Precipitated DNA
12
DNA Report Card™
EDVO-Kit #
119
Preparation of the DNA Report Card
22. Obtain a DNA Report Card™ and complete the identification information at the top of the card.
The Experiment
23. No sample is to be placed in the top circle labeled "Control (No
DNA)".
24. Use the transfer pipet from step 5
and 14 of the previous procedures
related to DNA precipitation:
EDVO-Kit
®
Slowly and gently transfer one drop
of the unstained DNA sample to the
designated circle on your DNA Report Card™. Allow the DNA sample
to dry.
25. Use the same pipet to slowly and
gently transfer one drop of the
stained DNA sample to the designated circle on your DNA Report
Card™. Allow the DNA sample to
dry.
26. Thoroughly wash the transfer pipet
with distilled water, or use a fresh
transfer pipet if you wish to repeat
steps 24 and 25 to assure that sufficient DNA is stored on your DNA
Report Card™.
27. After the samples have completely
dried, fold the DNA Report Card™
in half (top to bottom) and store in a
dry place.
# 119
DNA Report
Card™
Name: __________________
__________________________
Date:
__________________
Control
(No DNA)
Unstained
DNA
Methylene
Blue
Stained
DNA
Copyright © 2004, EDVOTEK, Inc.
DNA Report Card™
figure not drawn to
scale. Actual card
may appear slightly
different.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
13
119
Study Questions
1. Describe the appearance of the isolated DNA.
2. What do cell nuclei contain?
3. What are nucleotides?
5. What does cell lysis mean?
6. Why did the rubbing alcohol form a layer on top of the DNA solution?
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
The Experiment
4. How many chromosomes do humans have?
14
DNA Report Card™
EDVO-Kit #
119
xxx
Experiment Notes
EVT 007176K
EDVOTEK - The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
119
Instructor’s Guide
Class size, length of laboratory sessions, and availability of equipment are
factors which must be considered in the planning and the implementation
of this experiment with your students. These guidelines can be adapted
to fit your specific set of circumstances. If you do not find the answers to
your questions in this section, a variety of resources are continuously being
added to the EDVOTEK web site. In addition, Technical Service is available from 9:00 am to 6:00 pm, Eastern time zone. Call for help from our
knowledgeable technical staff at 1-800-EDVOTEK (1-800-338-6835).
E
O
DV
-TE
C H S E RV I C E
Technical Service
Department
1-800-EDVOTEK
ET
(1-800-338-6835)
Mo
Mon - Fri
9:00 am to 6:00 pm ET
FAX: (301) 340-0582
Web: www.edvotek.com
email: [email protected]
m
6p
n - Fri 9 am
Please have the following
information ready:
• Experiment number and title
• Kit lot number on box or tube
• Literature version number
(in lower right corner)
• Approximate purchase date
Online Ordering
now available
Visit our web site for information
about EDVOTEK’s complete line
of “hands-on” experiments for
biotechnology and biology education.
EDVOTEK - The Biotechnology Education Company®
1-800-EDVOTEK • www.edvotek.com
FAX: (301) 340-0582 • email: [email protected]
EVT 007176K
15
16
DNA Report Card™
EDVO-Kit #
119
Pre-Lab Preparations
• 1
Package of two sterile cotton-tipped applicators
• 3
Small transfer pipets
1. Add 200 µl of Tris
buffer to the tube of
protease. Allow the
material to hydrate
for a few minutes
and transfer the
entire amount back
ti the remaining Tris
buffer.
• 1 Calibrated transfer pipet
• 1 DNA Report Card
Each Student should receive:
Instructor’s Guide
• 600 µl Lysis buffer in clear tubes
with screw-on caps
Reagents to be Shared by Two Students:
Mix well and aliquot 100 µl for each
student pair. Be sure
to place the tubes
on ice until they are
needed.
• 100 µl Protease solution
• 200 µl NaCl solution in 0.5 ml
• 200 µl Methylene Blue solution
• 2 ml Ice cold alcohol (95% ethanol or
Isopropyl rubbing alcohol)
2. Aliquot 2 ml of Ethanol or Isopropyl
alcohol for each student pair. Place
on ice until needed.
3. Aliquot 200 µl of NaCl solution for
each student pair.
4. Aliquot 200 µl of Methylene Blue
solution for each student pair.
5. Cut DNA report cards™ and provide
one card for each student.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
DNA Report Card™
EDVO-Kit #
17
119
Inquiry-based Optional Extension Experiments
Encourage students to think of different parameters in the experiment
to alter and have them predict the outcome. Listed below are several
examples of optional activities for students to try.
Perform an experimental control with just the lysis buffer (cheek cells
are not added). Add the protease and NaCl and overlay the solution with alcohol. Note the differences between this control tube and
the sample tubes containing DNA.
•
Perform the DNA isolation and eliminate the protease, NaCl, and/or
alcohol from the isolation steps. What happens to the DNA sample?
•
After the addition of alcohol, isolate the DNA by spooling. DNA can
be recovered by dissolving in distilled water overnight and then prepared for agarose gel electrophoresis (materials for electrophoresis
not included).
•
Isolate the DNA and perform PCR on the DNA sample (materials for
PCR not included).
Expected Results
EDVO-Kit
®
# 119
DNA Report
Card™
Name: __________________
__________________________
Date:
__________________
Control
(No DNA)
The figure at left shows the DNA Report Card
before students perform the epxeriment. After completing the experiment, The Control
circle (top) on the DNA Report card™ will be
empty. The Unstained DNA circle(middle)
will appear slightly grayish white in color.
The stained DNA circle (bottom) will appear
blue.
Unstained
DNA
Methylene
Blue
Stained
DNA
Copyright © 2004, EDVOTEK, Inc.
DNA Report Card™
figure not drawn to
scale. Actual card
may appear slightly
different.
Duplication of this document, in conjunction with use of accompanying reagents, is permitted for classroom/laboratory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
Instructor’s Guide
•
18
DNA Report Card™
EDVO-Kit #
119
Study Questions and Answers
1. Describe the appearance of the isolated DNA.
Instructor’s Guide
DNA looks like a clear liquid gel. It turns white when it forms a precipitate.
2. What do cell nuclei contain?
The nucleus is an organelle present in human cells. It contains the
chromosomes that are rich in DNA.
3. What are nucleotides?
Nucleotides are the building blocks of DNA.
4. How many chromosomes do humans have?
There are 46 chromosomes (23 pairs) in human cells.
5. What does cell lysis mean?
Cell lysis is the disruption of a cell and cell nucleus with the release of
the contents.
6. Why did the rubbing alcohol form a layer on top of the DNA solution?
Alcohol has lower density than water and therefore will “float” above
the DNA solution.
document, in conjunction with use of accompanying reagents, is permitted for classroom/laboraDuplication of this
tory use only. This document, or any part, may not be reproduced or distributed for any other purpose without the
written consent of EDVOTEK, Inc. Copyright © 2004, 2006, EDVOTEK, Inc., all rights reserved
EVT 007176K
The Biotechnology Education Company® • 1-800-EDVOTEK • www.edvotek.com
Section V - Reactivity Data
Material Safety Data Sheet
EDVOTEK
®
IDENTITY (As Used on Label and List)
Section I
Emergency Telephone Number
Manufacturer's Name
EDVOTEK, Inc.
Telephone Number for information
Address (Number, Street, City, State, Zip Code)
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
Conditions to Avoid
Stable
Incompatibility
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Methylene Blue Plus ™
Unstable
Stability
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
(301) 251-5990
(301) 251-5990
X
Hazardous Decomposition or Byproducts
Toxic fumes of Carbon monoxide, Carbon dioxide, nitrogen oxides, sulfur oxides, hydrogen, chloride gas
Hazardous
Polymerization
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
OSHA PEL
ACGIH TLV
Route(s) of Entry:
Inhalation?
Health Hazards (Acute and Chronic)
Carcinogenicity:
Other Limits
Recommended
% (Optional)
Eyes: May cause eye irritation
Specific Gravity (H 0 = 1)
2
No data
Vapor Pressure (mm Hg.)
No data
Melting Point
No data
No data
Evaporation Rate
(Butyl Acetate = 1)
No data
Flammable Limits
No data available
LEL
UEL
No data
No data
Water spray, carbon dioxide, dry chemical powder, alcohol or polymer foam
Special Fire Fighting Procedures
Self contained breathing apparatus and protective clothing to prevent contact with skin and eyes
EDVOTEK
Emits toxid fumes under fire conditions
Sodium Chloride
Emergency Telephone Number
Manufacturer's Name
EDVOTEK, Inc.
Address (Number, Street, City, State, Zip Code)
Telephone Number for information
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
CAS # 7647-14-5
Boiling Point
OSHA PEL
No Data
No data
Vapor Pressure (mm Hg.)
Vapor Density (AIR = 1)
(301) 251-5990
(301) 251-5990
05/01/04
Respiratory Protection (Specify Type)
MIOSH/OSHA approved, SCBA
ACGIH TLV
Special
Local Exhaust
Ventilation
Protective Gloves
Other
Required
Mechanical (General)
Eye Protection
Rubber
Chem. safety goggles
Rubber boots
Unstable
Conditions to Avoid
X
Strong oxidizing agents, strong acids
Hazardous Decomposition or Byproducts
Hazardous
Polymerization
Conditions to Avoid
May Occur
X
Will Not Occur
Section VI - Health Hazard Data
Route(s) of Entry:
Inhalation?
Ingestion?
Skin?
Yes
Yes
Yes
Health Hazards (Acute and Chronic)
IARC Monographs?
NTP?
-------------------------- No data ---------------------------
OSHA Regulation?
Irritation
Other Limits
Recommended
% (Optional)
Medical Conditions Generally Aggravated by Exposure
Unknown
Emergency First Aid Procedures
Inhaled: Move to fresh air Eyes/Skin: Flush with water for 15 minutes
Ingestion: Wash mouth with water and call physician
Section VII - Precautions for Safe Handling and Use
Steps to be Taken in case Material is Released for Spilled
Specific Gravity (H 0 = 1)
2
2.165
No data
Melting Point
801°C
No data
Evaporation Rate
(Butyl Acetate = 1)
No data
Place in bag. Ventilate area and wash spill site
Waste Disposal Method
Observe federal, state, and local regulations
Precautions to be Taken in Handling and Storing
Avoid skin contact.
Other Precautions
None
Flammable Limits
LEL
UEL
--------Non-combustible----------
Use extinguishing media appropriate to surrounding fire conditions.
None
Section VIII - Control Measures
Signs and Symptoms of Exposure
Section IV - Physical/Chemical Characteristics
None
Precautions to be Taken in Handling and Storing
Carcinogenicity:
Clear liquid
Unusual Fire and Explosion Hazards
Mix material with a combustible solvent and burn in chemical
incinerator equipped with afterburner and scrubber. Check local and state regulations.
Incompatibility
Soluble
Special Fire Fighting Procedures
Waste Disposal Method
Stable
Section III - Physical/Chemical Characteristics
Extinguishing Media
Ventilate area and wash spill site
Stability
Signature of Preparer (optional)
Section II - Hazardous Ingredients/Identify Information
Flash Point (Method Used)
Treat symptomatically
Section V - Reactivity Data
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Section I
Appearance and Odor
No data available
Work/Hygienic Practices
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
IDENTITY (As Used on Label and List)
Solubility in Water
Emergency First Aid Procedures
Other Protective Clothing or Equipment
Material Safety Data Sheet
®
No data available
Medical Conditions Generally Aggravated by Exposure
None
Section IV - Physical/Chemical Characteristics
Unusual Fire and Explosion Hazards
OSHA Regulation?
Other Precautions
Blue solution, no odor
Extinguishing Media
Inhalation: Cyanosis
Keep tightly closed. Store in cool, dry place
Soluble - cold
Flash Point (Method Used)
IARC Monographs?
Yes
Steps to be Taken in case Material is Released for Spilled
No data
Appearance and Odor
Ingestion?
Yes
Section VII - Precautions for Safe Handling and Use
Boiling Point
Solubility in Water
Skin?
Yes
NTP?
No data available
Section III - Physical/Chemical Characteristics
Vapor Density (AIR = 1)
None
Meets criteria for proposed OSHA medical records rule PEREAC 47.30420.82
Methylene Blue
3.7 Bis (Dimethylamino) Phenothiazin 5 IUM Chloride
CAS # 61-73-4
X
Section VI - Health Hazard Data
Signs and Symptoms of Exposure
Section II - Hazardous Ingredients/Identify Information
Conditions to Avoid
May Occur
Will Not Occur
Skin: May cause skin irritation
07/01/03
Signature of Preparer (optional)
None
Strong oxidizing agents
Section VIII - Control Measures
Respiratory Protection (Specify Type)
Ventilation
Special
Local Exhaust
Mechanical (General)
Protective Gloves
Rubber
Other Protective Clothing or Equipment
Work/Hygienic Practices
Yes
Other
Eye Protection Chemical safety
Wear NIOSH/MSHA respirator
Do not ingest. Avoid contact with skin, eyes and clothing.
EDVOTEK
®
Emergency Telephone Number
Manufacturer's Name
EDVOTEK, Inc.
Address (Number, Street, City, State, Zip Code)
Telephone Number for information
Date Prepared
14676 Rothgeb Drive
Rockville, MD 20850
Conditions to Avoid
X
Stable
Incompatibility
Note: Blank spaces are not permitted. If any item is not
applicable, or no information is available, the space must
be marked to indicate that.
Protease
Unstable
Stability
May be used to comply with OSHA's Hazard Communication
Standard. 29 CFR 1910.1200 Standard must be consulted for
specific requirements.
IDENTITY (As Used on Label and List)
Section I
Section V - Reactivity Data
Material Safety Data Sheet
(301) 251-5990
(301) 251-5990
05/01/04
Hazardous Decomposition or Byproducts
Hazardous
Polymerization
Unknown
Conditions to Avoid
May Occur
None
X
Will Not Occur
Section VI - Health Hazard Data
Route(s) of Entry:
Inhalation?
Ingestion?
Skin?
Yes
Yes
Yes
May cause allergic & skin reactions. Irritating to mucous
membranes and upper respiratory tract.
Health Hazards (Acute and Chronic)
Carcinogenicity:
Signature of Preparer (optional)
None
Unknown
IARC Monographs?
NTP?
-------------------------- No data ---------------------------
OSHA Regulation?
Signs and Symptoms of Exposure
Section II - Hazardous Ingredients/Identify Information
Hazardous Components [Specific
Chemical Identity; Common Name(s)]
CAS # 9001-91-7
OSHA PEL
ACGIH TLV
Irritation
Other Limits
Recommended
% (Optional)
Vapor Pressure (mm Hg.)
Vapor Density (AIR = 1)
Solubility in Water
Appearance and Odor
Section VII - Precautions for Safe Handling and Use
Specific Gravity (H 0 = 1)
2
No data
No data
Melting Point
No data
No data
Evaporation Rate
(Butyl Acetate = 1)
No data
Flammable Limits
LEL
N.D.
UEL
N.D.
Special Fire Fighting Procedures
Wear SCBA and protective clothing to prevent contact with skin and eyes.
None
Dissolve or mix the material with a combustible solvent and burn in a chemical incinerator equipped
with afterburner and scrubber.
Precautions to be Taken in Handling and Storing
May cause sensitization by inhalation and skin contact.
Water spray, carbon dioxide, dry chemical powder, alcohol or polymer foam
Unusual Fire and Explosion Hazards
Waste Disposal Method
Avoid eye and skin contact and inhalation.
Powder, no odor
No data
Wear protective clothing, goggles, and gloves. Sweep up and place in bag and hold for disposal.
Avoid raising dust.
Other Precautions
Section IV - Physical/Chemical Characteristics
Extinguishing Media
Steps to be Taken in case Material is Released for Spilled
No data
Soluble
Flash Point (Method Used)
Unknown
Emergency First Aid Procedures
Skin/eye contact: Rinse with water. If inhaled, remove to fresh air
If swallowed, rinse mouth with water.
Section III - Physical/Chemical Characteristics
Boiling Point
Medical Conditions Generally Aggravated by Exposure
Section VIII - Control Measures
Respiratory Protection (Specify Type)
Local Exhaust
Ventilation
Mechanical (General)
Protective Gloves
No
Yes
Work/Hygienic Practices
Other
None
None
Eye Protection Chem. safety goggles
Chemical resistant
Other Protective Clothing or Equipment
Special
Suitable equipment to avoid eye/skin contact
Avoid contact