Maloof Lab:PCR stuff

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Room 2115
Section of Plant Biology
1002 Life Sciences, One Shields Ave.
University of California Davis
Davis, CA 95616


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Important: Label tubes with buffer name, date, your initials, and tube number

10x PCR buffer (see Promega PCR buffer) for 20 ml

500 mM KCl (10 ml 1M stock)
100 mM Tris/HCl (pH 9.0) (2 ml 1M stock)
15 mM MgCl2 (0.3 ml 1M stock)
1% Triton X (2 ml 10% Triton X) # Kazu and Liron changed 1ml into 2 ml (Oct 21, 2015)

Add sterile MilliQ up to 20 ml in total.

Note 1: Renew 10xPCR buffer every 1~2 months.

Taq polymerase

1 (current ('07) stock Taq) : 9 (storage buffer)

MgCl2 (25 mM)

Add 250 µl 1M MgCl2 to 10 ml dH2O

300 µl aliquots in 0.5 ml tubes.

dNTP (10mM total, 2.5 mM each)

Add equal amount of dATP, dTTP, dCTP, dGTP (100 mM, Fermentas dNTP Set #R0186 or Kappa dNTP set KK1007) and dilute into 10 mM

Make 100 µl aliquots.