Editing 3161 pET28b-GST-Myc-HA-Flag-V5 (section)

== Restriction Digestion Analysis of 3159&nbsp;<span style="color: rgb(0, 0, 0); font-family: Arial, Verdana, sans-serif; font-size: 12px; white-space: nowrap;">pMiniT2-1FRG-HA-VL-kappa</span> ==

by Paul, 3/16/2018

Nde1 + Xho1 digestion:&nbsp;1803 (vetor)&nbsp; and 3159 (insert)

[[File:1803 3159 Nde1 Xho1.JPG|RTENOTITLE]]

Left: vector&nbsp;1803

Right: 3159, there are too many bands, too complicated

[[File:3159 EcoR1.JPG|RTENOTITLE]]

Cut out EcoR1 band, middle ~ 1kb, purified with mini silica column, elute with 20 ul H2O

digest&nbsp;with Nde1 + Xho1

[[File:3159 1803 Nde1 Xho1.JPG|RTENOTITLE]]

3/17/2018, cut out the smallest fragment, notice 1803 vector is overdigested, can not be used

[[File:1803 3159 digestion.JPG|RTENOTITLE]]

digest vector 1803 with Nde1-Xho1 (right lane)

Date:3/19/2018

Miniprep of 3161

Restriction enzymes:NdeI+XhoI

Buffer:Fastdigest

TIme:30min

Result:combine the rest of them except#2 to do sequence

[[File:IMG 3623.JPG|RTENOTITLE]]

Date:3/20/2018

'''PCR screening of 3161'''

Lane1,2:length 1300bp

Primers:

820-His-HA-His-T;

T7-term(R)

Lane 3: length 294

Primers:pCold-seq-F;

T7-term(R)

[[File:IMG 3630.JPG|RTENOTITLE]]
[[Category:Cloning]]