Eniola Johnson Date: October 17,2022

ID#: 620156899

DNA Paternity Lab

PART B

1. Steps involved in processing DNA samples for paternity report

Introduction
DNA (deoxyribonucleic acid) is the double-stranded helix that is found within the

nucleus of cells within the body. DNA carries the genetic information needed to be tested. It

can be tested for several things. It can be used to identify genetic mutations. DNA may also

be used to assist in criminal investigations. Another application is when Bodies may have

been mutilated due to accidents, DNA profiling is needed here. Additionally, it is used to

establish a value for a child's paternity. Paternity tests can be very accurate almost 100%. It is

done using something known as an STR test. STR stands for short tandem repeats. These

repeats found on the gene are the basis used for paternity testing.

Step 1: Collection of the sample

This could be done in two ways, blood tests or a less invasive cheek swab. The

father and child will be in a medical facility and must consent to take the blood test or

swab. The blood may be taken from the arms of the family. The swab is taken from

the cheeks of the patients by the patients or a licensed practitioner. One swab must be

done for each cheek and each should last approximately 30 seconds.
Eniola Johnson Date: October 17,2022
ID#: 620156899

Step 2: Extracting the DNA

The extraction of the DNA will differ based on the sample, whether blood or

tissue. The cells must be lysed to obtain their internal contents. Lyse comes from the

word lysis which means to disintegrate. Lysing is a process by which the cell is

broken open and the DNA within is retrieved. It may be done thermally, chemically,

or even with the use of enzymes for example Proteinase K.

PCR amplification of multiple STR loci simultaneously, or multiplexing utilizes

different coloured fluorescent dyes

Step 3: Separating DNA

The DNA obtained may be contaminated or mixed around with debris and

chemicals that may have been used in the previous step. It must be isolated and

purified. DNA is then precipitated using sodium ions which will make the molecules

less water soluble and help with stability. Alcohol is added and causes the DNA to

precipitate out of the aqueous solution because it isn’t soluble in alcohol. Adding a

protease will degrade any DNA-associated proteins. Purification is the next step. The

DNA is rinsed with alcohol, specifically isopropanol or absolute ethanol, removing

the rest of the unwanted debris. Then it is dissolved in water for proper storage. The

most popular method of separation though is electrophoresis because DNA is

negatively charged. This uses an electric field to move molecules through a gel-like

substance or matrix. They migrate to the positive electrode and as smaller fragments

move faster; the larger ones move slower. They are arranged in order of size and the

bands formed are compared throughout the family.

Eniola Johnson Date: October 17,2022
ID#: 620156899

Step 4: Genotyping using PCR

After separation, the DNA is copied using the PCR technique.

PCR is a process that produces millions of copies of a selected DNA sequence. DNA

polymerases and 2 primers are needed as well as large amounts of the 4 DNA

nucleotides. The primers are needed as a starting point for the new strand so the DNA

polymerase can go to work. The DNA that has been stored is then heated to separate

the double strand into single strands. It is cooled and the primers begin to bind to their

complement sequences. This is done to give a small segment of the DNA that is

desired. It is then heated once more as the polymerases build new strands of the DNA

by attaching the nucleotides until the segment is filled. Now you have the specific

copied sequence needed for the study.

Step 5: Testing

These copies are tested for genetic markers. Loci which are the locations of

these markers house two alleles (alternative forms of a gene) that come from each

parent. A comparison must be made to the population database based on the

phenotype the individual displays. After the known genetic markers are located, they

are compared and matched between the child and parent/s. A paternity report is

prepared.
Eniola Johnson Date: October 17,2022
ID#: 620156899

2. How is this paternity report analyzed?

A paternity report is a DNA test utilized to determine the relationship between a

mother, an alleged father and their child but is not limited to this portion of the family.

On your paternity test, you’ll have the index, the probability of paternity along with

data results placed in a table and the test. At least 15 different DNA locations are

listed in the locus column and then compares the data with the same locations for

other tested parties. A child inherits one copy of the DNA segment from the mother

and one from the father known as alleles. The number associated with each allele

represents the length of the DNA segment where the man tested is considered the

biological father of the child tested. The possible father and the child must match one

allele at each genetic location, except in cases where a genetic mutation exists. When

there are mismatches at most locations then the result will be called an exclusion

meaning the man tested is not considered the biological father. If only one allele is

present at a single location, it means the two alleles from each parent are the same

length.

PI stands for paternity index. The PI column on the table depicts the strength of the

genetic match between the alleged father and child at that location. The higher the PI

the stronger the match. Each PI is multiplied together to obtain the combined paternity

index or CPI. This ratio indicates how many times more likely it is that the man tested

is the biological father than someone randomly selected from the population with the

same ethnicity. The higher the CPI the stronger the results. If a man is excluded as the

biological father, then the CPI is always 0. The probability of paternity is based on the

combined paternity index and is calculated using mathematical formula. If the man

tested is not excluded as the biological father, then the percentage of probability is
Eniola Johnson Date: October 17,2022
ID#: 620156899

99.9% or higher if he is excluded as the biological father, the probability of paternity

is 0%. The probability can never be 100% because each man from the child’s

phenotypical race would have to be tested as well.

PART B

1. Paternity Tables

CASE 1: PATERNITY TABLE FOR ALLEGED FATHER AND CHILD ONLY

Locus Child Alleged Shared Frequency Formula Paternity
Father Allele of Allele Index (PI)
CSF1PO 7,10 7 7 0.04286 1/2q 11.666
D5S818 8,12 8,13 8 0.05 1/4q 5.000
D7S820 9,10 9,10 9,10 0.15714, (p+q)/4pq 2.363
0.32381
TPOX 11 11 11 0.22488 1/q 4.447
D18S51 16,19 16,21 16 0.18889 1/4q 1.324

CASE 2: PATERNITY TABLE FOR ALLEGED FATHER AND CHILD ONLY

Locus Child Alleged Shared Frequency Formula Paternity
Father Allele of Allele Index
D21S11 31, 33.2 31, 35 31 0.09218 1/4q 2.712
D5S818 13 15 None 1 0 0.000
THO1 6, 8 6, 9 6 0.10952 1/4q 2.283
Vwa 16 18,20 None 1 0 0.000
D18S51 13,18 16,20 None 1 0 0.000
Eniola Johnson Date: October 17,2022
ID#: 620156899

2. Calculations

Combined PI= Product of all the PIs of the loci used

CPI for A =11.666 x 5.000 x 2.363 x 4.447 x 1.324

CPI for A = 811.542

Probability of paternity:

(CPI x 0.5)/ [(CPI x 0.5) +0.5] x 100

811.542 x 0.5)/ [(811.542 x 0.5) + 0.5] x 100

= 99.88% (The alleged father is not excluded as the biological father)

References (APA format)

Endeavor DNA. (2021, September 1). What is the process for DNA testing? Endeavor DNA.

Retrieved October 16, 2022, from https://endeavordna.com/blog/process-dna-test/

DNA Diagnostics Center. (2022, July 29). How to read and understand paternity-test

results. DDC. https://dnacenter.com/project/how-to-read-and-understand-paternity-

test-results/

AlphaBiolabs. (2022, March 21). What are DNA markers and how can they tell us who a

child’s father is? | AlphaBiolabs UK. AlphaBiolabs.

https://www.alphabiolabs.co.uk/blog/paternity-testing-explained-what-are-dna-

markers-and-how-can-they-tell-us-who-a-childs-father-is/#:~:text=If%20the%20man

%20is%20the

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Eniola Johnson Date: October 17,2022
ID#: 620156899