User:Stuart McKellar/Notebook/Bird Sex Testing/2013/01/04
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Reconciling information and thoughts after break
I did some reading and thinking and research during the xmas break. This is the result.
Finally found some housekeeping primers: 18S-F908 18S a AGCGAAAGCATTTGCCAAGA 401 This study (SVB) 18S-R1309 18S a AGTCTCGTTCGTTATCGGAATT This study (SVB)
Temps and new primers
P2/P8 primers work best at 60C, confirming my lab work. (from http://www.academia.edu/1460726/Sex_Identification_of_Some_Pet_Birds_by_Polymerase_Chain_Reaction-Based_Methods )
also, this paper suggests using some new primers and was getting results very similar to my ones. Going to try and order NP an MP primers. See this paper for what a positive and negative result looks like using P2/P8 primers.
NP primer: 5’-GAG AAA CTG TGC AAAACA G-3’
MP primer: 5’-AGT CAC TAT CAG ATC CGG AA-3’
Also this page suggests using 1μL of 10μM primer to give final concentration of 0.5μM in 20μL. Maybe I should start running the reactions at 20ul instead.
EDIT THESE PAGES TO SEE NICE FORMATTING