WET LAB DNA Barcoding: From Samples to Sequences - Northwest ...

WET LAB
CLASS SET
Procedure:
[Note: Work in groups of up to 4 students.]
1. Obtain ice in your ice container. PCR primers should be kept cold.
2. PCR tubes are very small and do not permit complicated labels. On a separate sheet of paper or in your lab
notebook, make a table like the one below (Table 2.1) to list each DNA sample purified in Part 1 by you and
your group members, and the number that you will use to label your PCR tubes. Your PCR tube should
also include your initials, such as “JT” in the examples below.
Table 2.1: DNA Samples and PCR Test Tube Labels:
PCR Tube Label
Species (or sample number) from which DNA was purified
*1 ____________________________________________________
*2 ____________________________________________________
*3 ____________________________________________________
*4 ____________________________________________________
* Be sure to include your initials in addition to the tube number.
Example:
JT1
Joe Ting’s Salmon DNA from Puget Sound, Washington
3. The PCR tubes are the small, 0.2 ml clear tubes provided by your teacher (See Figure 2.1). Each tube contains
a small white bead that includes the Taq DNA polymerase, deoxyribonucleotides (dNTPs), and buffer. Obtain up
to five tubes for your group: one for each student, and one for the negative control.
4. Label each PCR tube as you described in Table 2.1 (for example, “JT1” or “JT2”).
Using Bioinformatics: Genetic Research
5. Label one PCR tube with one of your group member’s initials and a “0” (for your group’s Negative Control, for
example, “JT0”).
6. Obtain a 1.7 ml microfuge tube for each member of your group and label this tube as you described in Table 2.1
(for example, “JT1” or “JT2”).
7. Obtain a 1.7 ml microfuge tube for your group and label it with the initials and “0” for your Negative Control
(as in Step #5 above).
8. Briefly “spin down” each tube of PCR primers using a microcentrifuge. This will force all of the liquid into the
bottom of the tubes to ensure accurate measurement with your micropipette.
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©Northwest Association for Biomedical Research—Updated October 2012