PCR Amplification of DNA: Difference between revisions

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+==SOP==
+*[[SOP - Electrophoresis]]
 ==Materials==
-*Primers – order from IDT prediluted to 100 mM in TE.  Make working solution of 1 uM (100x) in water with both sense and antisense primers combined (300uL water, 3uL of each primer)
+*Primers – order from IDT prediluted to 100 mM in TE.  Make working solution of 0.4uM-1 uM (100x) in water with both sense and antisense primers combined (300uL water, 3uL of each primer)
-*dNTPs – dilute to 2 mM each in water, make single use aliquots (50 uL aliquots... 10uL of each dNTP, 460uL water)
+*DreamTaq Green PCR Master mix - contains dNTPs, polymerase, salts, and buffer with loading dye  https://www.thermofisher.com/order/catalog/product/K1081?ICID=search-product#/K1081?ICID=search-product
-*Template – generally 1uL or less of a plasmid miniprep
+*RNAse-Free water - comes in DreamTaq kit
-*Polymerase – use Pfu Turbo for cloning and Taq for noncloning.  Use appropriate buffer.
 ==Protocol==