What does the intensity of your fragment indicate about the

-What does the intensity of your fragment indicate about the success of your PCR?

-What factors might increase the intensity of your fragment (include both pre-PCR and post-PCR mechanisms)

The full form of PCR is Polymerase Chain Reaction, which is a method for amplifying DNA.

1. The intensity of fragments which indicate the success of PCR are as follows:

Template :

plasmid DNA ~1-5 ng.
Genomic DNA ~ 100 ng

Primers:

50 -100 pmoles of every primer

Buffer

Magnesium chloride:

1-5 mM

deoxynucleotides:

50-200 µM of every dNTP

Taq polymerase:

0.5 to 1.0 unit.

The volume of the PCR reactions to 50 µL, or even 10 µL, to save on reagents.

2. The factors that increase the intensity of fragment are as follows:

If the template is not good then inspite of of good techniquie and reagents used the amplified products cannot be obtained.

· Primer design:

The number of primer design affecting the factors include:

Complementarity, specificity
Length
Base composition:
Matching primer Tms

Self complementarity, secondary structure, primer dimers