Download - DNA Fingerprinting Supplies from Iowa State Student Preparation DNA and Enzyme Preparation
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: 515/294-4629.
The electrophoresis equipment can be obtained by contacting the Area Education Agency.
DNA Fingerprinting
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)
DNA Fingerprinting
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 group8 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.
DNA Fingerprinting
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.
DNA Fingerprinting
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.
DNA Fingerprinting
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.
DNA Fingerprinting
DNA and Enzyme PrepStep 2
• Pipette 17 µl of a 0.025 µg/µl concentration of pBR322 DNA into the tube labeled "C."
DNA Fingerprinting
DNA and Enzyme PrepStep 3
• In the tube labeled "1" pipette 17 µl of 0.15 µg/µl concentration of lambda DNA.
DNA Fingerprinting
DNA and Enzyme PrepStep 4
• In the tube labeled "2" pipette 17 µl of 0.075 µg/µl concentration of Ad-2 DNA.
DNA Fingerprinting
DNA and Enzyme PrepStep 5
• In the tube labeled "3" pipette 17 µl of 0.025 mg/ml concentration of pBR322 DNA.
DNA Fingerprinting
DNA and Enzyme PrepStep 6
• In the tube labeled "4" pipette 17 µl of 0.025 µg/µl concentration of pUC19 DNA.
DNA Fingerprinting
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.
DNA Fingerprinting