Hello everyone!
It might be a silly question, but as a beginner (currently undergrad researcher) I am very confused about weather use qPCR genotyping or CAST-PCR. By browsing thermofisher's website, I've came across both types of assays for one of the SNPs we are studying at the lab.
However, practically, what would be the difference between use either of them?
The mutation we are studying is BRAF V600E (rs113488022 - chr7:140753336 (GRCh38.p13))
Thanks in advance!
qPCR is a general category that describes using a specialized machine to amplify DNA and measure the quantity that is formed during each step of the amplification. The customization is how the product is detected (fluorescent dye? labeled probes? etc.) and what specific targets, tissues, etc. you are using.
CAST-PCR is one particular type of qPCR. Ask someone in your lab to chat with you about it.
Hi Lilian,
qPCR is used for measuring quantity of your target (ie, number copies of genes or precise sequences for qPCR on DNA, and expression of a gene for RT-qPCR on RNA or cDNA), whereas the castPCR will only used to genotype a variant, one mutation.
castPCR means "Competitive allele-specific TaqMan PCR", one evolution for the old but reliable ARMS-PCR.
see this paper for more explanations:
https://www.nature.com/articles/srep18592
all the best
fred
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