Question about chemcion S7824,THX - (Dec/21/2006 )
hello , it's my frist post. 
sorry ,my English is too poor, i'm thinking how to express my question properly ! 
I do MSP for MONTHS! but ,have nothing but dimer! today ,i find the DNA after bisulfite treatment can get the right thing with the primer of actin(use for RT-pcr). if this told me that my
bisulfite treatment unsucessful ?? 
I use the KIT chenciom S7824
the question
1 the first step :add 7ul 3M NAOH in I ug DNA in 100ul. then the totle volume is 100ul or 107ul?
2 the next step:in the protocol 37c for 10min.but some tell me 50c for 10min, which one is correct?
3 put 20ul NAOH in bisulfite adjust pH=5--------but i use NAOH more than 100ul ?
longing for answer!!THX!!
Hi there,
the use of Actin suggests and would confirm that you are not getting 100% conversion of your DNA, this is normal, you will need to design primers that target fully converted DNA.
As for the use of the chemicon kit, I would follow the instructions to the letter.
Nick
the use of Actin suggests and would confirm that you are not getting 100% conversion of your DNA, this is normal, you will need to design primers that target fully converted DNA.
so ,if I shuold try again?my primers come from published papers , and it seems all researchs use them, should I change them ?
As for the use of the chemicon kit, I would follow the instructions to the letter.
sorry ,I don't understand what you said. my E is too poor
thX!! THX very much!!!
Nick