Editing Yijia Li 3 12 2018 (section)

== Date:3/13/2018 ==

'''Test of 3165miniprep'''

Digest enzyme:BsrGI

Time:30min

Buffer:fastdigest

'''Results:not good, got three bands'''

[[File:Test of 3165.JPG|RTENOTITLE]]

'''Repeat subclone of 3152'''

Restriction enzymes:BsrGI+NheI

Buffer:NEB2

Time:30min

'''Result:Vector digest is good, but insert is not good.'''

[[File:Repeat subclone of 3152.JPG|RTENOTITLE]]

'''Test of 3165 miniprep digestion'''

1:SalI+BsrGI NEB2

2:SalI+KpnI Fastdigest

3:BsrGI NEB2

Time:30min

[[File:Test3165.JPG|RTENOTITLE]]

'''Digest of Vector 3130'''

made for Gibson mix

BamHI+KpnI

'''result:good, cut out and purify'''

[[File:Gibsonmix of 3130digestion.JPG|RTENOTITLE]]

'''PCR of 947'''

1281 as template

F1:945F+947R

F2:947F+945R

Extension time:1.5min

Annealling:55oC

Cycles:25

Volumn:25ul

2XPCR MIx:Platinum mix HiFi SUpermix

'''Results:good,combine F1+F2 and do the next step'''

[[File:PCR of 947F1+F2.JPG|RTENOTITLE]]

'''Digest of 3009pMiniT'''

Enzymes:BsrGI+NheI

Time:1hour

Buffer:NEB2

[[File:3009pminitedigestion1.JPG|RTENOTITLE]]

'''Digest of 30090pMiniT'''

Enzymes:XhoI+NheI

Time:30min

Buffer:Fastfigest

[[File:3009pminit2digestion.JPG|RTENOTITLE]]