HOOSIER DADDY?
YOU ARE NOT THE FATHER!
Ward’s 36-8551
Paternity testing lab activity
Washington H.S.
Kasi Bolden
MATERIALS INCLUDED IN KIT
Agarose
 5x TBE running buffer
 DNA stain
 Amber bottle
 Gel staining trays
 Biohazard bag
 DNA samples(mother, child 2 alleged


fathers)
Quic kTime™ and a
TIFF (Unc ompres sed) dec ompres sor
are needed to see t his pic ture.
MATERIALS NEEDED
Agarose electrophoresis chambers
 Power supplies
 Water or microwave
 Micropipets with tips
 Apron, lab markers & goggles
 UV transilluminator
 UV goggles

OBJECTIVE
Learn the process of agarose gel electrophoresis
 Perform the electrophoresis procedure
 Identify the most probable DNA match between
two alleged fathers

PRIOR KNOWLEDGE
DNA fragment length
 DNA fingerprinting technology vs. paternity
testing
 DNA molecules found in a human cell

FUN FACTS ABOUT DNA
DNA testing was used to prove that Thomas
Jefferson was in fact the biological father of at
least on child of his slave, Sally Hemmings.
 Electrophoresis is derived from the Greek
electros, meaning electricity and phoros, meaning
“to carry across”
 Agarose is highly purified for of agar, which is
material extracted from seaweed.
 Gel electrophoresis issues in genetic testing ,
paternity analysis, forensics, conservation
biology, animal behavior, taxonomy and
evolutionary biology

FUN FACTS CONTINUED
The first time DNA was used as evidence in a
trial was in 1985.
 The first time DNA evidence actually sent
someone to jail was in 1988.
 If all of the DNA molecules found in a single
human cell were lined up end-to end, they would
reach a distance a of about two meters. Yet, all of
these molecules are packed into a nucleus 10
millionths of a meter in diameter

ANNETTE GORDON-REED DISCUSSES 'PASSING' AND THE
EFFECT IT HAD ON LIVES AND FAMILIES. THE INTERVIEW WAS
RECORDED IN OCTOBER 2008 AT MONTICELLO'S ROBERT H.
SMITH INTERNATIONAL CENTER FOR JEFFERSON STUDIES.

http://www.youtube.com/watch?v=CHWtkml0suc
&feature=PlayList&p=C63B8B73DBE0CB39&pl
aynext_from=PL&playnext=1&index=23
TIME
 Day

1
Casting gels and diluting buffer: 30 min.
 Day
2
Loading and running gel: 60 min.
 Staining and analyzing gel: 60min

SCENARIO

Two men claim to be the biological father of child.
The courts have obtained DNA samples from the
mother, child and the two alleged fathers. As the
director of a genetic testing laboratory, you will
analyze the sample using the DNA fingerprinting
technique to help settle the paternity dispute.
DAY 1:CASTING AN AGAROSE GEL
1.
2.
3.
4.
5.
6.
Add 280mL of distilled water to 70mL of 5X TBE
concentrated to obtain 350mL of 1X buffer.
Melt the agarose gel using a boiling water bath or a
microwave.
Place the comb on the far side of the casting trays.
Pour approximately 25 mL melted agarose into the tray,
until it reaches a depth of about 3 mm (NOTE: The 1X
TBE buffer will be added after the gel has formed.)
Allow the gel to solidify for approximately 20-30 minutes.
Do not disturb
After the gel solidified, carefully remove the comb.
DAY 2: LOAD GEL
1.
2.
3.
4.
Place the wells closer to the negative ( black)
electrode.
Add the 1X TBE buffer until the gel it
completely submerge.
Load each DNA sample; 10µL of each DNA
sample.
#1 Mother in , #2 Child, #3 alleged Father1 &
#4 alleged Father 2
DAY 2 PROCEDURE
5.
6.
7.
Allow gel to run for approx. 30mins @125V or
until the dye migrates towards the red
electrode. You should see bubbles.
Carefully remove the gel, on the casting tray,
from the electrophoresis chamber. Pour approx.
100mL warm dilute. (If the lab period does not
allow time to stain and destain the gel, place
the gel in a resealable bag and add 1-2mL of 1X
TBE buffer and refrigerate the gel until the
next lab period.)
Accurately sketch the bands you see on the
blank gel in the analysis section.
DISCUSSION
Would you modify this lab? If so how?
 What level of science would you do this lab?
 Could you use this as a cross curricular activity?
 Thumbs up or thumbs down

FINALE

Based on the autoradiogram the child will have
inherited bands from the mother and the alleged
father.