hello everybody, Iam performing qMSP for MTHFR gene.

On Real time PCR: Iam getting result with UnMethylation primer but I dont get any result with Methylation Primer.

On MSP : Iam getting bound with Methylation Primer but I dont get any bound with UnMethylation Primer.

we have one set primer and use methprimer for this design.

our samples: DNA of Washed sperm

Iam extracting DNA from sperm without kit.

Iam using zymo bisulfit kit for bisulfit treatment.

product size :  174bp

Whats problem????

can anyone help me ,please????

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