H*N
2 楼
Can anybody share you experience with Gibson assembly? Planning to do it,
but would like to hear your tips and potential pitfalls with the technique.
Thanks in advance.
but would like to hear your tips and potential pitfalls with the technique.
Thanks in advance.
H*N
5 楼
Thanks to wne and ankyin. How large was your final product? How many
fragments did you use?
fragments did you use?
M*g
7 楼
Just use NEB gibson assembly kit or their newer version NEBuilder hifi DNA
assembly kit. you can clone up to 6 fragments. Make sure use their
competent cells, since the assembly rxn many times is not compatible with
competent cells from other companies such as invitrogen or homemade cells.
you can use NEBuilder software to design your primers: http://nebuilder.neb.com/.
I have assembled 4 fragments and plasmids are normally 7-8 kb. if you have
questions, just call NEB tech support, and they are helpful. that's what i
normally did.
hope this helps.
assembly kit. you can clone up to 6 fragments. Make sure use their
competent cells, since the assembly rxn many times is not compatible with
competent cells from other companies such as invitrogen or homemade cells.
you can use NEBuilder software to design your primers: http://nebuilder.neb.com/.
I have assembled 4 fragments and plasmids are normally 7-8 kb. if you have
questions, just call NEB tech support, and they are helpful. that's what i
normally did.
hope this helps.
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