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Hi folks,
I have new RNAseq data assembled with Trinity, and now I am trying to use qPCR for validation. I blasted a gene of interest from another species against the RNAseq data for primer design, but I have got something like this in return.
comp82676_c0_seq1
comp82676_c0_seq2
comp82676_c0_seq3
comp82676_c0_seq4
comp82676_c0_seq5
All of them are best hits and have identical score and E value. They only have tiny gap differences when I aligned them.
May you explain what are diffrences between them and which one I shall use for primer design? I want a primer to amplify both gene of interest and the closest hit from the new RNAseq data.
Thank you very much for your time and help!
I have new RNAseq data assembled with Trinity, and now I am trying to use qPCR for validation. I blasted a gene of interest from another species against the RNAseq data for primer design, but I have got something like this in return.
comp82676_c0_seq1
comp82676_c0_seq2
comp82676_c0_seq3
comp82676_c0_seq4
comp82676_c0_seq5
All of them are best hits and have identical score and E value. They only have tiny gap differences when I aligned them.
May you explain what are diffrences between them and which one I shall use for primer design? I want a primer to amplify both gene of interest and the closest hit from the new RNAseq data.
Thank you very much for your time and help!