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Unformatted text preview: Page 1 of 2 Bi 1x, Spring 2009 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. The master mix you will be using today is Invitrogen Supermix II, which...
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- Fall '09
- 1m, 5 l, pcr master mix