DNA Fingerprinting

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DNA Fingerprinting Supplies from Iowa
State Student Preparation DNA and Enzyme
Preparation
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Obtaining Supplies (Iowa Teachers Only) Teachers
should request the following supplies (next
slide) at least a week in advance. Supplies
should be ordered by contacting Lori Miller by
phone in Iowa, 1/800-643-9504 or 515/294-9818, or
by Fax [phone]. The electrophoresis
equipment can be obtained by contacting the Area
Education Agency.
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Supply List 6 boxes of pipette tips (already
autoclaved) 34 µl of 0.025 µg/µl pBR322 per
class section (minimum of 68 ml per school) 17
µl of 0.075 µg/µl Ad-2 DNA per class section
(minimum of 34 ml per school) 17 µl of 0.15
µg/µl l DNA per class section (minimum of 34 ml
per school) 17 µl of 0.025 µg/µl pUC19 DNA per
class section (minimum of 34 ml per school) 3
µl Bgl 1 per class section (minimum of 6 ml per
school) 15 µl reaction buffer per class section
(minimum of 30 ml per school)
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Supply List (cont.) 144 µl Carolina Blu Gel
Stain per gel (minimum of 2 gels) 732 µl Carolina
Blu Buffer Stain per gel (minimum of 2 gels) 1
bottle Carolina Blu DNA Stain 0.7 g agarose per
gel (minimum of 2 gels) 70 ml 10X TBE per gel
(minimum of 2 gels) 40 ml blue migration dye for
each student group 8 microcentrifuge tubes for
each student group Fingerprinting instructions
for each student Please send the unused
pipette tips/boxes, 10X TBE bottles (rinsed out)
and Carolina Blu DNA Stain back to the Office of
Biotechnology when finished.
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Preparation of the Student Materials

• The supplies can best be provided to the class in
  groups of five students.
• The DNA samples should be refrigerated until
  class time.
• It is too expensive for the Office of
  Biotechnology to provide DNA for every student in
  a class. The office will provide enough DNA,
  restriction endonuclease, and reaction buffer for
  a minimum of two groups of five students or a
  maximum of one group of five students in every
  class section, whichever is greater.

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DNA Fingerprinting Preparation of the Student
Materials(cont.)

• For the other student groups, use distilled water
  to replace the DNA, restriction endonuclease, and
  reaction buffer.
• Every group of students should be provided with
  the blue migration dye.

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DNA Fingerprinting DNA and Enzyme Lab
Preparation
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DNA and Enzyme PrepStep 1

• For each group of students label 7-1.5 ml tubes
  "C,1,2,3,4, N and D" with a permanent felt pen.
• Tube "N" will contain the enzyme mixture.
• Tube "D" will contain the migration dye.

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DNA and Enzyme PrepStep 2

• Pipette 17 µl of a 0.025 µg/µl concentration of
  pBR322 DNA into the tube labeled "C."

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DNA and Enzyme PrepStep 3

• In the tube labeled "1" pipette 17 µl of 0.15
  µg/µl concentration of lambda DNA.

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DNA and Enzyme PrepStep 4

• In the tube labeled "2" pipette 17 µl of 0.075
  µg/µl concentration of Ad-2 DNA.

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DNA and Enzyme PrepStep 5

• In the tube labeled "3" pipette 17 µl of 0.025
  mg/ml concentration of pBR322 DNA.

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DNA and Enzyme PrepStep 6

• In the tube labeled "4" pipette 17 µl of 0.025
  µg/µl concentration of pUC19 DNA.

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DNA and Enzyme PrepStep 7

• In the microcentrifuge tube (1.5 ml) labeled "N,"
  pipette 15 µl of reaction buffer and 3 µl Bgl 1.

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DNA and Enzyme PrepStep 8

• In a microcentrifuge tube (1.5 ml) labeled "D,"
  pipette 40 µl of blue migration dye.

DNA Fingerprinting