maybe I will use DHPLC for screening for mutations in CDKL5 gene. The sistem I will use is "Wave 4500" from Transgenomic company. I am trying to find wich volume each injection has, so I could see for how many injection 5 liters of buffer could be used. If any of you use this system could you please tell me?
the second point is: theres is a specific/recomended size that my PCR amplification should have? I can't find any information about that... The tecnique works with any size?
And what about the specifications for the primers used? CG %? primer size? Is there any thing?
One last thing: The company says that the buffers, once that they are opened, only have 15 days of valid conditions. My lab co-workers say that we could use it for a month but no more... I have a lot of samples to test, but if I can't do it in one month? and if I open the buffers botles first I have to do the protocol optimization..what if I take to long in this? what if is a realy hard and time-spender optimization?
I am new in this "science life" and I already have so many "what ifs" that I am almost scared
thanks a lot for reading