Direct sequencing of Bisulphite PCR product - (Feb/25/2010 )
i have done bisulphite treatment and have got excellent MSP bands.i want to proceed bisulphite sequencing using BSP's.is there any specific protocol or steps when u have to perform direct sequencing ?
You will need different primers from MSP primers to amplify the modified DNA. Once you get PCR products, send them for sequencing. To get enough products for sequencing, most likely you have to do nested or two rounds of PCR. You can use either forward and reverse primer for sequencing, but reverse primer always works better than forward one.