Hi,

Has anyone tried to synthesize the sgRNA without using a kit? Can anyone help me with the protocol?

I found a Protocol on Benchling blog but I am not sure if it is going to work.

In this protocol they suggest to order these oligos :

5' - TAATACGACTCACTATAGGNNNNNNNNNNNNNNNNNNNNGTTTTAGAGCTAGAAATAGCAAGTTAAAATAG - 3'

5' - AAAAAAGCACCGACTCGGTGCCACTTTTTCAAGTTGATAACGGACTAGCCTATTTTAACTTGCTATTTCT - 3'

sgRNA fwd primer: GGATCCTAATACGACTCACTATAG

sgRNA rev primer: AAAAAAGCACCGACTCGG

I dont understand why the primers have the same sequence as oligos and not the complementary sequence of oligos.

Thank you and have a nice weekend everyone