Stool genomic DNA Fast Isolation Kit
(Spin-column)
A fast kit for the isolation of genomic DNA from soil/silt/feces
For laboratory research use only
DP4601
50 preps
Kit Content, Storage, and Stability
Content
Storage
50 preps
RT
50 ml
Buffer A
-20oC
750 ul
Buffer B
RT
5 ml
Buffer C
RT
25 ml
Protein Precipitation Buffer
RT
18 ml
Elution Buffer EB
RT
5 ml
Purification Column AC
RT
50
Collection Tube (2 ml)
RT
50
Extraction Buffer
All reagents are stable for 12 months when stored properly.
Note
Buffer B may form precipitate due to low storage temperature. If necessary, dissolve the
precipitation by 65°C incubation and then cool down to room temperature before use.
Ensure the bottles are tightly capped when not in use. It prevents evaporation, oxidation and pH
change of reagents.
All centrifugation steps can be performed at RT
Principle
In this kit, innovative extraction and lysis system allows rapid celllysis and inactivation of cellular
nucleases. DNA purity is greatly ensured by efficiently removing of debris, contaminants and humine
acids by a specially treated DNA spin-column. Innovative washing solution removes trace contaminants
and pure DNA is eluted. Purified DNA can be directly used in downstream applications.
Features
1. Rapid, DNA isolation under 60 min
2. Compatible, suitable for various soils, stool and other types of soil
Ecoli s.r.o., Studenohorská 12, 841 03 Bratislava 47, Slovenská republika, IČO: 35815906, IČ DPH: SK2020224668
tel: +421 2 64789336 fax: +421 2 64789040, mobil +421 903 514 184, e-mail: [email protected], www.ecoli.sk
3. High purity, Purified DNA typically has an A 260/A280 ratio between 1.7 and 1.9
4. Isolated DNA longer than 50 Kb and can be directly used for most downstream applications,
including PCR, Southern-blot, restriction digestion etc.
Procedure
1. Accurately weigh 0.3 - 0.5 g stool sample to a 1,5 ml microtube, add 1 ml Extraction Buffer and 5 μl
Buffer A, vortex 1-2 min, mix thoroughly and then place into 37oC water bath for 10 min. Mix
thoroughly every 2-3 min.
2. Add 100 μl Buffer B, vortex 1-2 min, mix thoroughly and then incubate at 65oC for 10 min. Mix
thoroughly every 2-3 min.
3. Centrifuge at 10,000 rpm for 10 min and harvest supernatant to a new 1.5 ml microtube.
4. Add 1/3 volume of Protein Precipitation Buffer and mix thoroughly.
5. Incubate on ice for 8 min, then centrifuge at 13,000rpm for 10 min and harvest supernatant.
6. PURIFICATION COLUMN TREATMENT: Add 500 μl buffer C in the middle of purification column
placed into collection tube, stay for 1 min, centrifuge 1 min at 10,000 rpm and discard the filtrate.
7. Place the treated Purification Column to a new 1,5 ml icrotube. Transfer the supernatant from step 5
into treated Purification Column, centrifuge 1 min at 2000 g and collect the filtrate (it contains DNA).
8. Accuratly estimate the volume of flow-through, add 0.6 times volume isopropanol, mix thoroughly
and centrifuge at 13,000 rpm for 10 min. Carefully remove supernatant, invert the column for 2 min
and air dry. If the precipitation is not clean enough, wash by 70% ethanol twice and air dry the pellet.
9. Add 30 μl Elution Buffer EB to dissolve the pellet.
DNA can be stored at 2-8oC for few days, or at -20oC for long term storage.
Ecoli s.r.o., Studenohorská 12, 841 03 Bratislava 47, Slovenská republika, IČO: 35815906, IČ DPH: SK2020224668
tel: +421 2 64789336 fax: +421 2 64789040, mobil +421 903 514 184, e-mail: [email protected], www.ecoli.sk