Discussion:
Polymerase chain reaction problem
I have using sterile technique, made up in an eppendorf tube the following:
Sterile water 15 ul,
primer 1 @ 30ng/ul,
primer 2 @ 30 ng/ul,
DNA Preparation (listeria or e.coli 5 ul),
Mastermix 25 ul.
I mixed the tubes and heated them, and carried out the elecrophoresis
for staining to obtain a photograph.
I need to draw a diagram to explain why the precise target sequence is not amplified until the third cycle of the PCR, also, how is the value of the annealing temperature is determined for a given P.C.R protocol.