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Bi1x_2009_W3_S1

Bi1x_2009_W3_S1 - Bi 1x Spring 2010 Week 3 Session 1 PCR...

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Page 1 of 2 Bi 1x, Spring 2010 Week 3, Session 1 PCR PCR In today’s lab, you will use PCR to amplify a region of the 16S ribosomal RNA gene in the genomic samples you extracted in week 2. Materials: Sterile, ultrapure PCR-grade water 16S forward primer (“F”) 16S reverse primer (“R”) 2X PCR Master Mix Your purified bacterial DNA from last week PCR tubes Special handling notes: - All components of the PCR reaction must be kept on ice at all times. - Use only filtered pipet tips and change tips every time to avoid contamination. - Never mix your reactions by vortexing; only mix by pipetting up and down or flicking the tube gently. About the master mix: A PCR master mix is a concentrated solution of DNA polymerase, dNTPs (deoxynucleotide triphosphates: dATP, dTTP, dCTP, dGTP), and ions or additives—all the components required for PCR except DNA template and primers. Master mixes are more convenient than using separate components.
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