>20bp Insertion Mutagenesis - How to?
I need to insert 21-30 bps. I inserted 1-2 nucleotides before by simply using primers with these extra bps but 30 bps sounded a little too much to do it with primers. How should I do it? I have these in mind:
1) I can do it with long primers (e.g. 70 bps) with insertion bps in the middle.
2) I can do it sequentially with shorter primers with 5-6 insertions at a time (prefer not to)
3) I can do it with a kit.
I haven't used a kit before and don't know how they work. I m lazy and preferred to ask here instead of reading bunch of different kits' protocols. To be honest I prefer not to buy a kit also.
What is your suggestion? Do you think I can do it with 1st option?
Thanks in advance,
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