Please allow me to ask a foolish question, why people alway do suquencing with bisulfite PCR , instead of MSP product. If we find the methylation by MSP, Why can't we do senquening directly ?
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#2
pcrman
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Posted 05 November 2004 - 03:25 PM
Sequencing MSP products is meaningless, but I do see in the literature people mistakenly did that.
I figure that people who sequence their MSP products wants to confirm if their MSP results are true or not. Actually, they cannot. No matter you sequence MSP or USP products, they are amplified with bias and the methylation status of CpG sites within the PCR product will never tell you the true picture. If you sequence MSP products, you will get much higher methylation level, USP lower methylation.
I figure that people who sequence their MSP products wants to confirm if their MSP results are true or not. Actually, they cannot. No matter you sequence MSP or USP products, they are amplified with bias and the methylation status of CpG sites within the PCR product will never tell you the true picture. If you sequence MSP products, you will get much higher methylation level, USP lower methylation.
#3
hslab
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Posted 06 November 2004 - 08:19 AM
but at least , if the MSP direct sequencing can't give you the true picture, it can tell you the methylated CpG do exist? but the ratio beteen the methylation over unmethylation remain unclear.
do you agree with me?
do you agree with me?
