Troubleshooting for gibson assmebly. No colonie?

Dharmakiriti Srin @Dharmakiriti_Srin

30 April 2020 0 2K Report

I trying clone for almost 2weeks through Gibson assembly?

quick my procedure:

vector : 5103bp conc = 22.2ng/ul

insert: 387bp conc = 58.6ng/ul

I was trying to ligate with total of 10ul. I went for 1:3 ratio:

vector : 51.6ng

insert: 11.7ng

master mix : 5ul + insert + vector = 10ul.

25min in ice after gibson

1min heat shock.

2min in ice

1hr incubation.

Where I am getting wrong. PLEASE HELP :( :( :(

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