Primers for qPCR
Posted 16 March 2011 - 10:59 PM
Very new to the field and just starting things.
I need to follow the gene expression in a plant using real time RT-PCR. Where do I really need to start? I.e, how will I design the primer for a plant whose sequence is not known and similar works have not been done on the plant (as far as I am aware)?
Pls advice, cos for now I am at a lost space! thanks and appreciate any assistance.
Posted 17 March 2011 - 06:28 AM
Another approach is to do a library of fragments, then hybridize the extracted DNA (colony blot) with a similar gene or probe. Identify the clone, sequene, and then develop a specific primer pair.
None of these are easy or simple.
Posted 17 March 2011 - 11:57 PM
Posted 18 March 2011 - 08:02 AM
And in my opinion, you should figure out the specific gene for expression analysis. Then carry on your design work.
I think the first strategy is more feasible and time-efficient