13f - Laboratory 13f: Extracting DNA from Human Cells for...

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Laboratory 13f: Extracting DNA from Human Cells for PCR and Sequencing Background With a few precautions, DNA can be extracted from cells for PCR or DNA sequencing in much the same way as DNA is extracted from bacteria cells. Cells may be burst open and proteins precipitated out of solution. Ions that are released from the cell can interfere with Taq polymerase activity and inhibit the PCR reaction. To remove the ions out of the DNA sample, a step is added using the chelating agent, Chelex® (Bio-Rad Laboratories) chelating beads. Chelating agents bind ions and remove them from solution. Although some ions are needed in PCR, the technician adds the desired ions at the correct concentration when they are needed. Purpose To isolate human DNA from cheek cells for PCR and DNA-sequencing analysis. Procedure Part I: Reagent Preparation 1. For a team of four, prepare 50 mL of 0.9% NaCl solution. In your notebook, record the calculation for this preparation and include a drawing of how it is prepared. 2.
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This note was uploaded on 12/06/2011 for the course ENG 100 taught by Professor Posh during the Summer '11 term at San Mateo Colleges.

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