couldnt ligate 1.7kb into t vector

[ramanisubi]

hai

i was wondering if anyone suggest how to ligate a 1.7kb insert into T vector.
coz, ligating 1.7kb into 2.9kb vector will be reasonable?
the proportion i took vector: insert is 1:3 , is that right
for ligating higher length inserts, should we increase the vector concentration?


i tried 6 times, colonies were there, but none of them were positive clone

please help


-ramanisubi

[fatima]

Hi
The ratio 1:3 should be ok for that if you wish you can use 1:5 or6 but what you should verify is the polymarase you are using allow you to have a A since you are using a T vector. If You are using a T vector from a kit use the control to verify your ligation.
Good luck

[wafaamom]

the T vector has a specific conditions to get the insert in ? then ! but what is difference about T and any vector? and what is the meaning and the importance of the ratios here?