What is Hot Start PCR?
Hot Start PCR is a technique that inhibits Hot Start Taq polymerase activity or the incorporation of modified dNTPs during reaction set up until a heat activation step occurs. Hot…
How to do ARMS-PCR?
The process of the ARMS PCR is simple and very effective. No harmful radiolabelling is involved in the ARMS allele-specific PCR. The procedure of ARMS PCR is divided into 4…
What is ARMS-PCR or allele-specific PCR?
The allele-specific PCR also called as an ARMS- PCR (amplification refractory mutation system) or PASA (PCR amplification of specific alleles) or AS-PCR used to detect the SNPs. More specifically, it…
Role of nested PCR in microbial identification
The nested qPCR or the nested RT-PCR gives great power to this technique which can be a helpful method for the phylogenetic analysis and identification of different pathogens. The technique…
What is nested PCR?
In the nested PCR, the specificity of the PCR reaction is enhanced by reducing the non-specific binding with the help of the two sets of primer. The specificity is the…
Protocol for inverse PCR
In the beginning, before doing any wet lab work, design the primers complementary to the know DNA regions using the computational software. The detail information of how to design primer…
Inverse PCR
Here, the unknown DNA sequence adjacent to the known template DNA is amplified using the primers which amplify the flanking (unknown) region near the know DNA template. Quite difficult to…
Maxam-Gilbert Sequencing: What Was It, and Why
In the mid-1970s, two methods were developed for directly sequencing DNA: the Maxam-Gilbert sequencing (or chemical sequencing) method and the Sanger chain-termination method. Indeed, in 1980, both Walter Gilbert and Frederick…
Clone Library Screening
The result of any cloning experiment that begins with total DNA from a specific source is a library of clones. Thus cloning mRNA through a cDNA intermediate generates a cDNA library that represents the…
Prokaryotic Expression Vectors
The first vector we will consider is the pUC family of vectors Figure 3.4.1: pUC vector Although it is not typically used for the expression of recombinant proteins, it has all the necessary…
