THE PROFILING OF DNA ON FIRED
CARTRIDGE CASES AND PIPE
BOMB FRAGMENTS
Haylee De-Arne Bechaz
THIS THESIS IS SUBMITTED IN PARTIAL FULFILMENT OF THE
REQUIREMENTS FOR THE AWARD OF BACHELOR OF APPLIED
SCIENCE (HONOURS) AT THE UNIVERSITY OF CANBERRA
December 2001
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A w t m l r n Mce
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To figM crime and win
research centre
ABSTRACT
It is well documented that the rate of detection of identifiable fingerprints on fired cartridge
cases is low. Similarly, the detection of fingerprints on pipe bombs would be made
difficult due to the high temperatures and gases that are generated. Not to mention the
actual fragmentation of the pipe bomb components. Mitochondria1 DNA has been
recovered from fired cartridge cases, however it is not known whether nuclear DNA can be
recovered and profiled from a fired cartridge case.
This project aimed to determine the likelihood of sufficient nuclear DNA surviving on a
fired cartridge case or a detonated pipe bomb to enable profiling for forensic purposes.
The experiments carried out were designed to address the aim and examine the factors that
may affect the profiling of DNA from fired cartridge cases and pipe bomb fragments.
These factors include potential inhibitors of DNA that might be found at the crime scene,
DNA recovery and extraction techniques and transference of trace DNA.
The results indicate that nuclear DNA can be recovered and profiled from fired cartridge
cases if a significant quantity of DNA is present before firing. Significantly more DNA was
recovered from the fired aluminium cartridge cases compared with the fired brass cartridge
cases. Further studies are required to establish whether this result was a true trend or a
result of chance alone. More DNA was profiled for the pipe bombs containing propellant
than black powder, even though the pipe bombs containing propellant had a higher degree
of hgmentation. This may have been a result of DNA inhibition.
This research was a preliminary study with a low number of repeats. This was due to a
number of limitations. Further research is required in all areas with an emphasis on a
higher number of repeats.
ACKNOWLEDGEMENTS
I would like to express my sincere gratitude to my supervisors, Ian McNaught, Chris
Lennard, Julie Sutton and Paul McFawn. Thank you for all your time and valuable advice.
This year has been a wonderful experience and I can't thank you all enough.
Thanks are due to the Gadi Research Centre at the University of Canberra (UC) for their
assistance and support during the year.
Thank you to the AFP Bomb Squad for taking the time out of their busy schedule to help
me with my project. It was truly appreciated.
A big thank you must also go to the biology team at the Australian Federal Police (AFP)
Forensic Services Centre, Weston for putting up with me during this year. In particular,
Elizabeth Brooks for her constant encouragement and great sense of humour, which helped
me on a daily basis. Thanks to Eric also for taking the time out to take photos of my
cartridges under the SEM.
I also must thank Josephine Dachs who had to put up with me every day. I could not have
got through this year without your friendship. You helped me believe in myself, thank you
so much.
To my boyfriend, Matt, thank you for your positive energy and your way of always seeing
the bigger picture.
To my family, thank you for all your love, support and encouragement throughout the
years. I love you all very much.
TABLE OF CONTENTS
Page
..
ABSTRACT.. ...........................................................................................ii
...
STATEMENT OF ORIGINALITY. ..................................................................111
ACKNOWLEDGEMENTS...........................................................................
iv
...
LIST OF TABLES...................................................................................viii
LIST OF FIGURES....................................................................................ix
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Chapter 1 INTRODUCTION
1.1
Literature review.. ..................................................................
.2
1.2
Limitations..........................................................................
-14
-
Chapter 2 MATERIALS & METHODS
2.1
General Procedures..
...............................................................I6
2.1.1 DNA Swabbing.............................................................16
2.1.2 Trace DNA Extraction.....................................................17
2.1.3 Amplification...............................................................20
2.1.4 Capillary Electrophoresis.................................................22
2.2
.........................................................................24
Experiments
2.2.1 Dilutions......................................................................24
2.2.2 Inhibitors....................................................................
.26
2.2.3 Method of recovery and extraction.......................................28
2.2.4 Examination of fired aluminium and brass cartridge cases under a
SEM...........................................................................
30
2.2.5 Different surfaces...........................................................
31
2.2.6 Shedability....................................................................33
2.2.7 Handling of unfired cartridges............................................34
2.2.8 Detonation of pipe bombs ..................................................35
2.2.9 Firing of cartridges..........................................................39
Chapter 3 .
RESULTS
3.1
3.2
3.3
3.4
..............................................................42
DNA ratio............................................................................42
Limitations of the statistical analysis involved...............................43
Experimental results...............................................................44
Calculation of results
3.4.1 Dilutions....................................................................
-44
3.4.2 Inhibitors.....................................................................45
3.4.3 Method of recovery and extraction.......................................48
3.4.4 Examination of fired aluminium and brass cartridge cases
under a SEM.................................................................
50
3.4.5 Different surfaces...........................................................53
3.4.6 Shedability...................................................................55
3.4.7 Handling of udired cartridges............................................56
3.4.8 Detonation of pipe bombs.................................................57
3.4.9 Firing of cartridges..........................................................63
Chapter 4 .
DISCUSSION
4.1
4.2
..............................................70
Discussion of Experiments........................................................73
Discussion of General Procedures
4.2.1 Dilutions....................................................................-73
4.2.2 Inhibitors..................................................................... 74
4.2.3
Method of recovery and extraction........................................76
4.2.4
Examination of fired aluminium and brass cartridge cases under a
SEM...........................................................................
78
4.2.5
Different surfaces...........................................................79
4.2.6
81
Shedabilty....................................................................
4.2.7 Handling of unfired cartridges............................................82
4.2.8
Detonation of pipe bombs .................................................83
4.2.9
Firing of cartridges..........................................................86
CONCLUSIONS & FURTHER RESEACRH........................................88
APPENDIX..........................................................................................98
Quantification using the Rotorgene 'Real Time' 2000 machine...............99
Appendix 1a ........................................................................
100
Appendix 1b .......................................................................
-101
Appendix 1c ........................................................................-104
Appendix 1d .......................................................................-106
Appendix l e .......................................................................-108
Appendix If .......................................................................
-109
Appendix 1g ........................................................................
110
vii
LIST OF TABLES
2.1
The dilutions that were used for experiment 2.2.1 .......................................24
2.2
The volume of solutions used for inhibitor experiment.................................27
2.3
Technique and volume of solutions used for recovery and extraction
experiment....................................................................................
.29
3.1
The recovery of DNA from different square areas of washed hands from four
individuals......................................................................................55
3.2
The DNA ratios for DNA recovered from each fired cartridge case...................63
LIST OF FIGURES
1.1
Loading a fixed and detachable magazine pistol ..........................................3
1.2
A diagram of a DNA allelic ladder profile ................................................12
2.1
Diagram of a Microcon 100.................................................................17
2.2a
Photo of black powder being poured into the pipe bomb ................................37
2.2b
Photo of the hole dug for the pipe bombs ..................................................37
2 . 2 ~ Photo of the hole dug covered with wire mesh netting and several sandbags........37
2.2d
Photo of a pipe bomb fragment .............................................................37
2.3
Loading a single cartridge into the ejection port of the firearm.........................40
3.1
The average DNA ratio calculated for each DNA quantity.............................44
3.2
The decrease in average DNA ratio with increasing volumes of black
powder ..........................................................................................
45
3.3
The effect of increasing quantities of dirt supernatant on the amplification of
DNA ............................................................................................
46
3.4
The effect of increasing quantities of propellant on the amplification of DNA......46
3.5
Comparison of recovery techniques of DNA from glass microscope slide..........48
3.6
The difference in average DNA ratio between extraction techniques .................49
3.7
SEM photo of the surface of a fired brass cartridge case (320 x magnification)....50
3.8
SEM photo of the surface of a fired aluminium cartridge case
(300x magnification).........................................................................50
3.9
SEM photo of the surface of a fired brass cartridge case (3000 x magnification) ...51
3.10
SEM photo of the surface of a fired aluminium cartridge case
(3000 x magnification) .......................................................................51
3.1 1
Comparison of the recovery of 1 ng DNA from three different surfaces.............53
3.12
Comparison of the recovery of 10 ng DNA fi-om three different surfaces............54
3.13
The recovery of DNA from brass and aluminium cartridges after two
individuals handled them for approximately 10 seconds................................56
The pipe and endcaps recovered from one of the detonated pipe bombs containing
black powder ...................................................................................
57
The fragments that were collected from the second detonated pipe bomb containing
black powder ..................................................................................-57
The level of fragmentation experienced with the detonation of a pipe bomb
..
contammg propellant ..........................................................................
58
The pipe bomb fragments collected after detonation using propellant .................58
The level of fragmentation of one of the pipe bombs containing Powergel ..........59
The tiny fragments that were collected after the detonation of a pipe bomb
..
contamng Powergel ..........................................................................
59
61
The recovery of DNA between the explosives used .....................................
The average DNA ratio calculated from two different explosive charges............62
The recovery of 40 ng DNA fiom brass and aluminium fired cartridge cases.......64
The recovery of 239 ng DNA from brass and aluminium fired cartridge cases......65
The results from the fired brass cartridge cases containing 40 ng DNA ..............66
The results from the fired aluminium cartridge cases containing 40 ng DNA .......67
The results from the fired brass cartridge cases containing 239 ng DNA .............67
Categorises the results from the fired aluminium cartridge cases containing 239 ng
DNA ...........................................................................................
-68