PCR conditions

Recommended amplification conditions for the original SSLPs are as follows: 
20 ul reactions containing 
1-10 ng genomic DNA
5 picomoles of each primer 
200uM deoxyribonucleotides 
50 mM KCl
10 mM Tris-Cl pH 9 
0.01% gelatin 
0.1% Triton X-100
2 units of Taq polymerase. 

The final concentration of magnesium chloride was usually 2 mM. 

Standard cycling conditions were: 
94C for 15 seconds, 55C for 15 seconds and 72C for 30 seconds, repeated 40 times. 

Note:
Locus ca72 requires an annealing temperature of 61C 

nga112 requires 3 mM magnesium.

The PCR conditions for the subsequent SSLPs are
as follows, except where noted. 

N.B. The following conditions generally work with the original SSLPs.

PCR-The program (30) should run for 30 cycles and should be:  
Denature at 94 degrees for 15 seconds, 
Annealing at 55 degrees for 15 seconds, 
Extend at 72 degrees for 30 seconds. 

The program H30 is the same as 30 except the primers require an initial start at 94 degrees for three minutes. 

The gels will either be a 3% BRL agarose gel or a 3% metaphor agarose gel in 1X TAE.