T-DNA insertion screening - T-DNA (Jun/30/2008 )
Hi there,
I got two T-DNA insertion lines. One from SALK and one from SAIL. I am not certain, whether those lines are homozygous for the insertion or heterozygous. So far I was only able to identify 6 heterozygous out of 20 samples.
My question is: How many DNA-samples do I have to screen in order to be sure, that there are really no homozygous T-DNA lines?
My second question concerns the SAIL lines. Unlike the SALK lines, there are three different T-DNA specific primers (LB1, LB2, LB3). Which of those do I have to take? How do I find out, which of those primers applies to my T-DNA lines?
I got two T-DNA insertion lines. One from SALK and one from SAIL. I am not certain, whether those lines are homozygous for the insertion or heterozygous. So far I was only able to identify 6 heterozygous out of 20 samples.
My question is: How many DNA-samples do I have to screen in order to be sure, that there are really no homozygous T-DNA lines?
My second question concerns the SAIL lines. Unlike the SALK lines, there are three different T-DNA specific primers (LB1, LB2, LB3). Which of those do I have to take? How do I find out, which of those primers applies to my T-DNA lines?
My professor gave me 6 TDNA insertion lines from FLAG. I don't had the experience of growing them but I tried and still i'm getting some problems. So would you please give me the protocol for growing TDNA knock out mutants. I first grew them through tissue culture in MS media and then transplant into pots. But transplant survival rate is very low. I don't know where i'm wrong? I need your help
Hello
I will be making transgenic rice lines using agrobacterium mediated transformation. Would you please send me detail protocol. I need each and every steps even minor steps too. thank you