Editing Yijia Li 3 12 2018 (section)

== Date:3/14/2018 ==

'''947 F3 PCR'''

F3:1,2

Negative Control:3

PCR Mix:Platinum

Primers:945F+945R

'''Result:Have bands, cut the top one and do the next step.'''

[[File:947F3PCR.JPG|RTENOTITLE]]

'''Miniprep digestion_3161_3168'''

3161:1,2 NdeI+XhoI Buffer:Cutsmart

3168:3 BsrGI+NheI

Buffer:NEB 2

Time:40min

'''Result:seems good, do sequencing'''

[[File:Miniprep of 3161 3168.JPG|RTENOTITLE]]

'''Miniprep digestion of 3131_3161_3167'''

Name length Enzymen Buffer

3131:1-4 342 BssHII+XhoI Cutsmart

3161:5-8 225 NdeI+XhoI Cutsmart

3167:9-12 397 BsrGI+NheI NEB 2

Time:40min

[[File:Miniprepof 3131 3167 3161.JPG|RTENOTITLE]]

'''Redo miniprep of 3131_3161_3167'''

3131:1-4 BssHII+XhoI 342 Cutsmart

3161:5-8 NdeI+XhoI 225 Cutsmart

3167:9-12 BsrGI+NheI 397 NEB 2

Time:40min

[[File:Redo miniprep of 3131 3161 3167.JPG|RTENOTITLE]]

'''miniprep digestion 3021'''

3021pMiniT2:BssHII+XbaI 374

3021:BssHII+XbaI 374 Cutsmart

3165:SalI+BsrGI 702 NEB 3

Time:30min

[[File:Redominiprep diegstion 3021.JPG|RTENOTITLE]]

'''Subclone of 945'''

1:1727 XbaI+NcoI 173&nbsp; Cutsmart

2:1834 XbaI+NcoI 173&nbsp; Cutsmart

3:945pMiniT XbaI+NcoI 1326&nbsp; Cutsmart

4:3009pMiniT BsrGI+NheI 371&nbsp; &nbsp;NEB 2

Time:30min

'''Result:1727 can see band as vector, 945pminiT2 can see band as insert, cut out and combine'''

[[File:Subclone of 945.JPG|RTENOTITLE]]

'''Redo Subclone of 3167'''

Vector:3163pCEH0

Insert:3027pMiniT2

BsrGI+KpnI

Buffer:NEB 2

Time:30min

'''Result:Cut out and combine'''

[[File:Redo Subclone 3167.JPG|RTENOTITLE]]