Hi :)
How should I design primers for (-)ssRNA samples from Influenza A and hRSV viruses?
I need to select the protein, locate the gene sequence, and then use it to design primers. Should I retrieve the cDNA sequence from NCBI to work with?
The term 'cds' in FASTA means that it is the sequence without introns. Is it okay to use this type?
if you have any guides, videos or documents you would like to share with me, I would be very grateful.
Thanks
Designing specific primers for reverse transcription (RT) involves several key considerations to ensure efficient and specific amplification of the target RNA. These considerations include target selection, primer length, GC content, Tm calculation, avoidance of self-complementarity, specificity, positioning, optional incorporation of a T7 promoter for IVT, quality control, and optional modifications. Following these guidelines can help optimize primer design and ensure successful RT-PCR amplification of the target RNA.
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