Rensselaer/16 September 2008

Setting up primers for PCR

Added Nuclease free water to dried primers to obtain a 100 micromolar concentration stock.

e.g. 30.5 nm(1/350 uL) = 100 pm/uL

primers for PCR are to amplify the YbjX promotor and the CopA promoter. Two forms of the 3' YbjX has been included due to a possibility of one of the primers containing a ribosomal binding site. These primers were set in 1.5 mL centrifuge tubes and diluted 10x to a total volume of 200 uL.

e.g. Vol(primer) + Vol(water) = 200 uL

V1 uL x 100 umolar = 200 uL x 10 umolar V1 = 20 uL of primer to add to tube. 200 - 20 = 180 uL of water to add to column (add water first).

Diluted primers and stock were set in -20 Celsius freezer.