My solution to this.Re: cloning problem# Biology - 生物学
z*h
1 楼
I just did it 1.5 month ago.
Let me explain my ways,
For double disgested vector, using agrose to separate the DNA fragment and
using QIAGEN gel kit to get it.
For mutated DNA fragment, you should design the primer containing restriction
enzyme cutting size plus several NT to ensure the resricting enzymes can work
efficiently. Check this information from Appendiex of NEB catalogue.After
digestion, using DNA concentrator kit to remove enzyme, salt and small end of
DNA fragment.
Mix your vector pl
Let me explain my ways,
For double disgested vector, using agrose to separate the DNA fragment and
using QIAGEN gel kit to get it.
For mutated DNA fragment, you should design the primer containing restriction
enzyme cutting size plus several NT to ensure the resricting enzymes can work
efficiently. Check this information from Appendiex of NEB catalogue.After
digestion, using DNA concentrator kit to remove enzyme, salt and small end of
DNA fragment.
Mix your vector pl