Question

1. Compare PCR to qPCR, pros and cons

2. What must be considered when making PCR primers associated with annealing conditions? How to minimize errors when designing primers to anneal to the DNA template

Answer 1

PCR is the term used in biotechnology for the production of multiple copies of DNA.

The main advantage of qPCR is that it requires 1000 folds less template DNA for the reaction to proceed.

qPCR detects,characterizes and quantifies the nucleic acid for numerous applications.  

qPCR combines nucleic acid amplification and detection in a single step, whereas pcr quantifies the product at theend of pcr cycles

PCR qualitatively detects nucleic acids whereas, qPCR offers more sensitive quantitative analysis of the sample.

2. Length and primer's composition affects the PCR annealing temperatures. A melting range of 52-58C is a good starting range when designing primers. lengthy primers and primers with high G and C content have higher melting temperatures. For best amplification the primers should be 17-24 bases long. Shorter primers efficiently anneal to the target DNA.

To avoid errors during annealing to the DNA template, the primers should be specific to the target DNA. the DNA sample should be pure without any contamination that might inhibit Taq. Primer dimer formation can be reduced by adjusting the concentrations of primer and dna as recommended