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Can human 18S primer, probe set for qPCR be used for Rhesus and Cynomolgus sampl

qPCR primer probe real time pcr

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#1 witioni

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Posted 28 August 2012 - 01:54 PM

Going to create an 18s pr/pb set for use with a human, rhesus, and cynomolgus assay.

Found a set from literature but not too sure if it will pick up rhesus and cynomolgus. Nothing shows on blast.

Cannot use ABI set because we want to modify the primer set a bit and, of course, ABI will not release the sequence.

Will this primer probe set developed for human 18s work on Rhesus and Cynomolgus? I would believe so considering the gene is evolutionarily slow to change and the close relationship between all three species.

Thoughts?

Thanks in advance!

#2 bob1

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Posted 28 August 2012 - 02:03 PM

You should be able to primer blast against the rhesus and cynomolgus genomes, assuming they have been published.

#3 witioni

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Posted 28 August 2012 - 02:13 PM

Did a blast but it did not pick up rhesus... which seems unusual to me considering the close relation and how evolutionarily conserved 18s is, no? Cyno is not published.

#4 bob1

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Posted 28 August 2012 - 03:07 PM

Did you try to blast it specifically against the rhesus genome (under the database choose the "others" option and nr/nt from the dropdown list. Then type in macaca in the organism field. It is more specific and may help find the site you want. If you choose Blastn under the program selection, that may help tell you how close the sites are.





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