Professional Documents
Culture Documents
TECHNIQUES
Pranab Chatterjee
1. METHYLATED DNA
2. SHORTCOMINGS OF RFLP
3. SHORTCOMINGS OF PCR
4. SHORTCOMINGS OF SOUTHERN
BLOT
5. MT-DNA IN MATERNALLY
RELATED
6. STANDARDISATION
7. SIMILARITY AMONG RELATIONS
B.1) METHYLATED DNA
– Contamination
– Technical faults can be magnified
tremendously to alter the result
completely
– Selection of wrong primers in
shallow gene pool areas or in
people of same family, etc.
A robot used in DNA profiling adds solution and stirs DNA
samples from tissues
B.4) SHORTCOMINGS OF SOUTHERN
BLOT
1. Cost efficiency
2. Comparative analysis
3. To determine acceptability
4. Stutter Bands
5. Shallow gene pool
6. Jeffrey Gafoor Case
7. The O. J. Simpson case
8. The Dr. Scheenberger case
C.1) COST EFFICIENCY
Now, P(C1A)=P(CUA)
P(A)
FromBayes’ Theorem,
P(A1C)=P(CUA) P(A)=P(C1A)
P(A)………1
P(A) P(C) P(C)
The probability that the accused is guilty is
hinged upon two facts:
Accused is guilty and has DNA +ve i.e.
P(CUA)
Accused is not guilty and has DNA +ve
i.e. P(CUA’)
In all other cases, the accused has DNA –ve
and is ruled out.
Thus, probability that the accused has
DNA+ve is—
P(C)= P(CUA)+P(CUA’)=P(CUA)P(A) +
P(CUA’) P(A’)
P(A) P(A’)
From Bayes’ Theorem,
P(C)= P(C1A) P(A) + P(C1A’) P(A’)
Also P(A)= 1—P(A’)
Therefore, from equation 1 we have putting P(C):
P(A1C)=P(C1A) P(A)= P(C1A) P(A) .
P(C) P(C1A) P(A) + P(C1A’)
P(A’)
= P(A) .
P(A) + P(C1A’) P(A’)
P(C1A)
Let p= P(A) and r= P(C1A’) , then
P(C1A)
P(A1C)= p .= p .
p + r.P(A’) p + r(1-P(A))
So,
P(A1C)= p/ {p + r(1-p)}
This is known as Matthew’s
confessional probability.
THE STATISTICAL CONSIDERATIONS:
Now, for a DNA positive Prior Odds defendant is
accused to be guilty, Assumption
innocent given the
frequency of the
Probabilty that the accused is of Guilt DNA profile, and
incorporating prior
Guilty given that DNA is +ve> assumptions of
probability that the accused is guilt
innocent given that DNA is 0.000 1
+ve
For the accused to be guilty, 0.001 1 in 1,002
then, the following probability 0.10 1 in 111,112
has to be satisfied:
0.50 1 in 1,000,0001
P(A1C)>P(A)
This, in turn implies that r<1 0.90 1 in 9,000,001
The calculation of probable
0.99 1 in 99,000,001
guilt is tallied alongside:
Thus we see that the 50%
assumption of guilt tallies with the
accepted frequency of the DNA
profile in the population.
THUS IT IS STATISTICALLY
SAMPLE COLLECTIONS
Sperm:
1oo µl semen taken, centrifuged at
5000rpmx 5 min
Supernatant discarded, TNE buffer
added, Dithioerithritol and SDS
added ProteinaseK added before
overnight incubation at 37o C
Follow from step d of fresh whole
blood sequence
RECOMMENDATIONS
Tissues:
Tissues frozen in liquid nitrogen and
homogenised
TNE buffer added, followed by SDS and
Proteinase K. overnight incubation at 55o
C
Follow from step d of fresh whole blood
sequence
RECOMMENDATIONS
Hair:
Hair roots are taken and crushed gently
TNE buffer with SDS and Proteinase K added
and incubated overnight at 55oC
Follow from step d of fresh whole blood
sequence
RECOMMENDATIONS
QUESTIONS?
pranabchatterjee@hotmail.com