WET LAB DNA Barcoding: From Samples to Sequences - Northwest ...
WET LAB DNA Barcoding: From Samples to Sequences - Northwest ... WET LAB DNA Barcoding: From Samples to Sequences - Northwest ...
WET LAB CLASS SET 21. Add 30 μl of your purified PCR product to your sequencing sample tube by gently but firmly pushing the pipette tip through the gasket on the lid of the tube (see Figure 4.3). After putting your sample in the tube, carefully pull your pipette tip out of the tube, leaving the gasket in place. 22. Give your DNA sequencing tube containing your sample to your instructor. Figure 4.3: Pipetting the Sample into the Operon Sequencing Tube by Gently Forcing the Pipette Tip through the Gasket. Using Bioinformatics: Genetic Research 366 ©Northwest Association for Biomedical Research—Updated October 2012
WET LAB KEY Lab 1: DNA Purification for DNA Barcoding Teacher Answer Key 1. On a separate sheet of paper or in your lab notebook, describe what happened to the cell membrane. Where did it go? The cell membrane was digested or broken down by the detergents in the lysis solution. Pieces of the membrane are floating around in the buffer in the microfuge tube with the DNA. 2. On a separate sheet of paper or in your lab notebook, describe what your sample looks like. If students began with a sample of the appropriate size (i.e., not too big), the cells should be completely lysed, and the samples should be clear, indicating that all of the membranes have been dissolved. 3. What did you do in this laboratory experiment and why? Students purified DNA from sample X to perform DNA barcoding. Students may go into greater detail about the specific steps of the protocol: breaking open the cells, dissolving the membranes, and removing the DNA from the rest of the cell debris. The important thing is for students to understand that genetic research starts with DNA, and we must have DNA purified away from the other cellular components for subsequent experiments. 4. What skills did you learn or practice? This question is designed to help students identify laboratory skills that they can list on a resume and/or college application. • Handling samples • Pipetting • Microcentrifugation • DNA purification using spin columns 5. List at least three types of samples from which you could purify DNA, based on what you have learned about tissues in your classes. What type of cells or tissues could you not purify DNA from? This question is designed to help students realize that DNA is present in, and can be isolated from, many different types of cells/tissues. Common samples include: • Muscle • Blood • Saliva • Shaft of bird feathers (analogous to the root of a mammalian hair) Wet Lab – DNA Barcoding: From Samples to Sequences 367 ©Northwest Association for Biomedical Research—Updated October 2012
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<strong>WET</strong> <strong>LAB</strong><br />
CLASS SET<br />
21. Add 30 μl of your purified PCR product <strong>to</strong> your<br />
sequencing sample tube by gently but firmly pushing<br />
the pipette tip through the gasket on the lid of<br />
the tube (see Figure 4.3). After putting your sample in<br />
the tube, carefully pull your pipette tip out of the tube,<br />
leaving the gasket in place.<br />
22. Give your <strong>DNA</strong> sequencing tube containing your sample<br />
<strong>to</strong> your instruc<strong>to</strong>r.<br />
Figure 4.3: Pipetting the Sample in<strong>to</strong> the Operon<br />
Sequencing Tube by Gently Forcing the Pipette Tip<br />
through the Gasket.<br />
Using Bioinformatics: Genetic Research<br />
366<br />
©<strong>Northwest</strong> Association for Biomedical Research—Updated Oc<strong>to</strong>ber 2012
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