3 replies to this topic

[InduS]

Hi guys,
I'm new to this forum
I'm very much new to this field of methylation studies. Can anyone tell me which is best method to study the demethylation; methylation specific PCR or Real time PCR? Can you please justify this?
By the way excellent forum

[pcrman]

If you are not going to do high throughput mapping, the commonly used candidate gene methylation study methods are bisulfite modification based PCR methods which involve first treating DNA with busulfite to convert unmehtylated Cs to Us, then PCR amplification of the converted DNA. There are two PCR methods: MSP and BSP. If you choose MSP, you can use qPCR or regular PCR. In BSP, you amplify the template to get enough DNA for direct sequencing or sequencing after cloning.

[InduS]

If you are not going to do high throughput mapping, the commonly used candidate gene methylation study methods are bisulfite modification based PCR methods which involve first treating DNA with busulfite to convert unmehtylated Cs to Us, then PCR amplification of the converted DNA. There are two PCR methods: MSP and BSP. If you choose MSP, you can use qPCR or regular PCR. In BSP, you amplify the template to get enough DNA for direct sequencing or sequencing after cloning.

Please tell me what this high throughput mapping is? I've googled it. But not satisfied with the answer.

[pcrman]

High throughput mapping means to assess DNA methylation on a genome wide scale or thousands of CpG islands.