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genotyping (SNPs)

qPCR internal control

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#1 SF_HK



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Posted 22 January 2020 - 06:26 PM



Im doing genotyping for a few SNPs by Taqman probe qPCR. Im wondering what I can use as internal control?


My samples are DNA and the program in my QS6 is genotyping. Im running water as my negative control. 


For sybr green qPCR, my internal control would be any house keeping gene e.g. GAPDH.


Im wondering internal control in qPCR for SNP genotyping?


Thnak you



#2 OldCloner



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Posted 26 January 2020 - 07:54 AM



Unlike RNA expression-level qPCR, you don't need a separate endogenous gene (like GAPDH) as a control.  SNP determinations are not quantification assays, they are plus/minus assays for each allele. It is recommended that you include, in addition to your no-template negative control, a positive control DNA sample with a known genotype,  If possible include 3 separate samples, one for homozygous allele 1, one for homozygous allele 2, and one known heterozygote.  These are desirable, but not essential to the running of the assay. This is confirmed by the Taqman SNP manual in Table 12, page 17. (URL below)





Good luck!

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