Hi everyone,
We are trying to amplify the COI sequence from mosquito gDNA samples using the following universal barcoding primer pairs:
Folmer et al., Molecular Marine Bioliology & Biotechnology, 1994
LCO 1490 5′-ggt caa caa atc ata aag ata ttg g-3′ (25 bases)
HCO 2198 5′-taa act tca ggg tga cca aaa aat ca-3′ (26 bases)
However, we are not able to get amplicons. we have checked the PCR reagents and the gDNA extracts with other primer pairs. they are working fine.
Primers also seem to be ok, based on the synthesis report and run on agarose gels and performance of other primers synthesized during the same batch.
Any help regarding any other primer pair for the same purpose, or any troubleshooting will be highly appreciated.
Thank you in advance.
S Datta
https://www.diva-portal.org/smash/get/diva2:657046/FULLTEXT01.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4863023/pdf/ECE3-6-3001.pdf
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4282734/pdf/13071_2014_Article_569.pdf
Dear Dr El-Hindi,
Thank you for the very useful links. We will try these works.
regards,
S Datta
Thanks for sharing. This is very significant paper for testing all important mosquitoes. specially primer information.
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