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)*1 * Add sterile MilliQ up to 20 ml in total. * Note 1. Kazu and Liron changed 1ml into 2 ml (Oct 21, 2015) * Note 2: Renew 10xPCR buffer every 1~2 months. Homemade Taq polymerase ------------------ * see protocol [attachment:19]. * Current working TAQ is 1:5 diluted (May 2017 by Lynn). 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. * Current one (as of Aug 2017) is Gold Bioscience (www.goldbio.com, 1ml of 10 mM each). Dilute four times and store them in your box (do not return diluted one to lab stock).