Editing 2437-pT7A30-Tat-NLS-Cas9

&nbsp;'''Date:&nbsp;''' '''2015-April'''

'''Clone ID: 2437'''

'''By: Xin Xu'''

'''Results''':&nbsp;

Clone TAT into vector 2332 pT7A30-Cas9 by NdeI +NcoI

PCR 1708 using

PCR on 1708 pT7A3-HTNCre<br/>[[File:2437 PCR.JPG|201x139px|RTENOTITLE]]

Digested the vector 2332 pT7A30-Cas9 by NdeI +NcoI<br/>[[File:2437 vector.jpg|RTENOTITLE]]

Ligation, transformation, screening:

[[File:2437 screening.JPG|RTENOTITLE]] &nbsp;

PCR screening:

The primers used for screening was inside the gene: 2437-Tat-NLS-Nco1-R;2437-Tat-NLS-Nde1-F

All the clonies have the right PCR Product.



PCR screening again, using one primer was on the vector Pet-seq-F2 and one inside the gene 2437-Tat-NLS-Nco1-R



[[File:2437 PCR SCREENING with pet.JPG|230x138px|RTENOTITLE]]

The colonies have a faint product. Also the -ve control also have the product. contaminated?

sent No3 to sequence.