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Introduction
It is common to hear reports in the media of crimes that have been solved with the
assistance of DNA analysis (Brown, 2006). If enough blood, semen or tissue is found
at a crime scene, forensic laboratories can determine the blood type or tissue type by
using antibodies to detect specific cell surface proteins (Campbell et al, 2006). This
method unfortunately requires large amounts of fresh samples and is not a strong
source of evidence as several people have the same blood or tissue type and thus it
only narrows down a few suspects (Campbell et al, 2006). DNA testing has a high
level of certainty as DNA sequencing is unique for each individual except for
To obtain a profile a Short Tandem Repeat (STR) needs to be generated from blood or
other samples (Bustamante et al, 2007). Profiling makes use of the STRs which are
short sequences, 1-13 nucleotides long that are repeated several times in tandem array.
In DNA profiling, the alleles of a selected number of different STRs are determined
(Brown, 2006).When restriction enzymes are used to cut STRs and the results are
DNA-modifying enzymes that cut DNA at specific sequences produce DNA segments
which are called restriction endonucleases or restriction enzymes (Knox et al., 2005).
The fragments that are formed when DNA is digested by restriction enzymes can be
joined together in new combinations using DNA ligase to create recombinant DNA
(Knox et al, 2005). DNA fragments of different sizes are separated by gel
molecules in a mixture under an applied electric field (Lodish et al., 2004). The
dissolved molecules in an electric field move at a speed determined by their charge to
mass ratio. Many proteins or nucleic acids have very similar charge to mass ratio and
various gels rather than in a liquid solution (Lodish et al., 2004). In this experiment,
we will only use restriction enzymes and agarose gel electrophoresis techniques for
Refer to Biology 1004 Molecular Biology practical manual, “Week 8, DNA Profiling
Experiment was carried out as per the lab manual, with the exception in the procedure
where 5µL samples of DNA were used in step 1 opposed to the outlined 2µL and a
Aim
To investigate how DNA profiling may be used to assist in solving a crime using
Results
The DNA samples were run through gel electrophoresis where fragments of different
1 2 3 4 5 6 7 8
Figure 1. Gel slide of DNA bands extracted from the three suspects found under UV light. The
amplified fragments are visualized on agarose gel. Lane 1: DNA Ladder. Lane 2: Blank. Lane 3:
Suspect X. Lane 4: Suspect Y. Lane 5: Blank. Lane 6: Suspect Z. Lane 7: Evidence sample. Lane
8: Blank.
Using the information from the gel photo (see figure 1), an initial comparison can be
closely matched with the evidence, but at this point further calculations are required to
An accurate measurement of the distances between each fragment can then be taken
from figure 1 as the number and sizes of the DNA bands are known and the distance
3.5
size of fragment (log bp)
2.5
y = -0.0652x + 4.7011
2
1.5
0 5 10 15 20 25 30 35 40 45 50
distance travelled (mm)
Figure 2. Standard curve for the DNA ladder of the fragments (in log measurements) in relation to
the distance travelled of each band. The added trend line and its equation provides the basis for
further calculations of DNA samples.
Running a sample of a DNA ladder in the gel slide also assists in the analysis. The
distance travelled by each fragment was measured from figure 1 and a predetermined
ladder was provided by the demonstrator to help identify the size of fragments. Using
this information a standard curve for the ladder was produced (see figure 2). From this
curve, a trend line was added which produced an equation which provides the basis
for further calculations in the DNA samples of the suspects for this case.
The measurements of samples X, Y, Z and E were taken from the wells, and using this
information and the equation provided from figure 2, an accurate fragment size could
Sample calculations:
Distance travelled = 30 mm
Therefore for size in bp, take natural log to the power of 10.
102.75 = 556.03 bp
Using the calculations above, it is possible to further investigate the fragment sizes in
each sample and evaluate whether it is probable that the evidence presented matches
2000
1800
1600
Fragment Size (bp)
1400 Sample X
Sample Y
1200
Sample Z
1000
Evidence
800
600
400
200
0
1 2 3 4 5
Fragment Sample
Figure 3. The size of each fragment (bp) compared to other DNA samples from the three suspects.
Again, sample Y appears to be closely matched to the evidence as their lines appear to
have the most similar trends as depicted in figure 3 (see appendix for further details).
Although sample Z follows similar trends initially, sample Y coincides with the
evidence at fragment points 4 and 5 in figure 3, setting it apart from the other samples
tested.
Discussion and Conclusion
as the distance travelled and its fragment size are not so closely matched to the values
presented in the evidence. Sample Z appears closely matched to the evidence sample
in both figures 1 and 3, but the fragment size that is presented after measurements and
calculations appears to be slightly larger than that of the evidence. Nevertheless, this
sample still represents a likely suspect involved at the crime scene. Sample Y showed
similar trends in Figures 1 and 3 to the evidence provided for this case and further
comparison across groups in the class confirms the trends found across all samples
would have given a more precise account of the distance travelled by the DNA bands.
loading and running the agarose gel. Errors to this process (or other steps in the
method) would cause an unclear scan on the agaraose gel making a proper analysis of
At a crime scene, the biological evidence found, such as blood, semen hairs etc,
would be unintentionally left behind and the quantities would be very small. DNA
fingerprinting might be used in such cases with the assistance of polymerase chain
reaction (PCR), where the STRs are amplified before testing, thus the problem of
In criminal cases, analysis of DNA fingerprints requires more than a test through
evaluates the chances of another likely candidate within a population of having the
exact genetic information. It would not prove an individual’s guilt but rather
strengthen a case, along with other DNA evidence, relating to a person’s involvement
in a crime.
In conclusion the sample that most closely matches the evidence presented is that of
Sample Y, the boyfriend of the deceased. This is due the similarities in fragment size
and trends calculated from this analysis. It should be noted that the presence of this
individuals DNA at the scene of the crime does not necessarily imply that he was
involved in the murder, but rather his presence at that location at some point in time.
References
Cummings, M.R (2003) Human Heredity: Principles and Issues, 6th Edition. Pp 323-
325. Brooks/Cole, Pacific Grove, CA. Knox, et al. (2005) Biology: an Australian
Lodish, H., Berk, A., et al. (2004) Molecular Cell Biology, pp 87-89, W.H Freeman
Appendix
Nat. Size
Sample X Distance trav (mm) Log (bp)
Fragment 1 30 2.75 556.03
Fragment 2 31 2.68 478.52
Fragment 3 33 2.55 354.41
Fragment 4 35 2.42 262.48
Nat. Size
Sample Y Distance trav (mm) Log (bp)
Fragment 1 22 3.27 1847.99
Fragment 2 23.5 3.17 1475.37
Fragment 3 25 3.07 1177.88
Fragment 4 28 2.88 750.76
Fragment 5 35 2.42 262.48
Nat. Size
Sample Z Distance trav (mm) Log (bp)
Fragment 1 23 3.2 1590.38
Fragment 2 25 3.07 1177.88
Fragment 3 27 2.94 872.37
Fragment 4 30 2.75 556.03
Fragment 5 31 2.68 478.52
Nat. Size
Sample E Distance trav (mm) Log (bp)
Fragment 1 22.5 3.23 1714.35
Fragment 2 24 3.14 1368.67
Fragment 3 26 3.01 1013.68
Fragment 4 28 2.88 750.76
Fragment 5 35 2.42 262.48
Appendix questions
(see attached)