What is the best clonning method (TA, blunt end, etc...) for clonning small PCR fragments (20-30bp) to later sequence and blast for species identification?
RT-PCR reaction is generating Ct values for non-specific targets. Need to identify these targets in an effort to design better PCR primers or to explain the non-specific amplification. Sequence homology is the no brainer reason but I need to identify the organism that is homologous to my target sequence.
Please help if possible. Clonning efforts are wearing thin.