Primer Design..need very basic advice.
Posted 01 November 2010 - 07:07 AM
Can someone please take me through a "fake" example on how to do this manually? And how do you design the PCR cycling based on what primers you have?
--Indebted to those who reply
Posted 01 November 2010 - 03:03 PM
Why is this happening?
Posted 02 November 2010 - 11:56 AM
Therefore, to quantitate the methylation on strand A, the forward sequence will show C/T peaks at the C position of CG dinucleotides. In the opposite direction, the sequence will show G/A peaks in the G position of CG dinucleotides. This is because you are only looking at the methylation in the C position of the original A strand.I think this is where you are getting confused - the C position in the reverse strand is always going to show C as it corresponds to the original G. It is the G position in the reverse which needs to be measured, as this is the position which corresonds to the C in the orignal A strand.
Confused about the Bolded part? Can someone explain in further details...thank you.