Mad researcher

Hello,

So, i got through my PhD interview. But the professor said he is 95% sure/interested in hiring me? He has two more interviews to take and he will let me know after that.

I was wondering what does 95% mean as in how sure is he or how interested is he.

I am running a multiplex PCR with sDY (sex determining primer) and 18s (control) primer to determine the sex of juvenile rainbow trout.

I am using 1.25 uL of PCR buffer (sigma), 250 micro molar of dNTP, 0.25 uL of Jump start Taq DNA and 0.4 microl molar of (sDY primer both F and R) and 0.1 micro molar of 18s Primer (both F and R). i take 40 ng of DNA and its a 12.5 uL reaction.

The problem i am facing is that i should get a single band around 400 bp if it is a female and double bands around 200-210 bp if it is a male. But now i am getting a faint (lighter) band which shouldn't appear in case of the female.

PCR CYCLE:

Initial denaturation at 94 degrees for 1 minute, denaturation for 60 sec at 94°C; 35 amplification cycles of 94°C for 30 sec, 60°C for 30 sec and 72°C for 30 sec; and a final extension of 3 min at 72°C

--------------------------------------------------------------

What should i change so as to avoid the double band in case of female? I have already considered reducing the amount of DNA (to 30 ng).

A motivation letter is the same as statement of purpose (SOP = for US universities).

So what should i write in the body of the email?

Is anyone using Gel red for gel electrophoresis?

I needed to ask something about this product so if anyone has any experience in using this then please let me know.

Moved and merged so as to keep recent discussion in one place. Bob

you can try for another post-doc position elsewhere or join an industry as a post doc