( ria strains indicated. ...)Optimizing DNA Amplification Protocols using the Eppendorf Mastercycler

ria strains indicated. A 123456L B Result:

Evidence of 2 different large rRNA operons in the genome of Heterorhabditis was not provided until the primer concentration, the dNTP concentration and the amount of polymerase had been optimized.

A: 1: Heterorhabditis HSH1
2: Heterorhabditis HSH2
3: Heterorhabditis HB1
C: Positive control
L: 100-bp ladder Standard PCR conditions prior to optimization
Annealing temperature : 58C
Volume : 50 l
dNTPs : 200 M each
Primers : 2 M each
Taq polymerase : 2 U B: 1,4: Heterorhabditis HSH1
2,5: Heterorhabditis HSH2
3,6: Heterorhabditis HB1
L: 100-bp ladder Optimized PCR conditions
Annealing temperature : 54C (1-3) and 58C (4-6)
Volume : 50 l
dNTPs : 20 M each
Primers : 0.2 M each
Taq polymerase : 0.5 U

Polymerase concentration

The concentration of Taq polymerase is a critical factor in determining the stringency of a PCR.
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