Product information

Easy PCR Kit

traditional PCR Kit including all reagents

A5200

Introduction
The-Easy PCR kit contains all components for PCR amplification of specific DNA regions. Just add your own template DNA and primers for the particular experiment. The kit contains Taq DNA polymerase, PCR nucleotide mix, buffer and a magnesium chloride solution. The positive control (high molecular weight DNA) and control primers provide the confidence in PCR control reactions. The kit is available in 2 sizes - for 100 and 500 reactions in 50 microliters. Control DNA and primers are sufficient for 10 reactions in 50 microliters. The size of the amplicon synthesized with the use of control primers and control DNA is 316 bp. The-Easy PCR kit contains 100 reactions 10X Taq DNA polymerase buffer without MgCl2 A5200,0100A 160 mM (NH4)2SO4, 650 mM Tris HCl (pH 8.8 at 25C), 0.1% Tween-20 dNTP mix (10 mM each) A5200,0100B Taq DNA polymerase (5 units/l) A5200,0100C 25 mM MgCl2 A5200,0100D Nuclease-free water A5200,0100E Control DNA A5200,0100F Control primer mixture A5200,0100G Protocol One PCR reaction (50 l) contains: 10X buffer (without MgCl2) dNTPs mixture MgCl2 Taq DNA polymerase Control DNA Primer1+2 mixture H2O 5 l 1.8 l 8 l 0.5 l 1 - 2 l 2 l up to 50 l 1 ml 200 l 50 l 1 ml 2 x 1 ml 22 l 22 l 500 reactions A5200,0500A A5200,0500B A5200,0500D A5200,0500C A5200,0500E A5200,0500F A5200,0500G

Recommended Mastermix (50 l / reaction): Example: 1 Reaction 10 Reactions 10X buffer (without MgCl2) 5 l 50 dNTPs mixture 1.8 l 18 MgCl2 8 l 80 Taq DNA polymerase 0.5 l 5 H2O 28 l 280 total volume 43,3 l 433 Add to each tube either Control DNA & Control Primers or your own Template/Primers. H2O fill up to 50 l

A total volume of 25 l may be sufficient. You may safe reagents, when you prepare a mastermix for several test. Recommended thermocycling: Initial heating 2 minutes 30 cycles 10 sec 25 sec 25 sec final fill-in 10 minutes

94C 94C 55C 72C 72C

Time mentioned for each step doesn't include the heating-up and cooling-down time, only the time at the referred temperature is considered. Since thermocyclers of various producers doesn't show uniform heating and cooling, the cycling characteristics may differ for other PCR machines. So, ideally, the temperature, time of each step and MgCl2 concentration should be optimized empirically for each machine and for the particular pair of primers.