Editing 3165 pEF1-HTLV-1IFH-HA-VH-mIgG2b (section)

== Redo ==

'''3165 PCR 1st'''

F1:3028 as template

Primer:3165F1+3165R1

length440bp

extenstion time:1:30

F2:3129 as template

Primer:3165F2+3165R2

Buffer:Platinum

length:165bp

Extension time:15bp

[[File:3165PCRF1+F2.JPG|RTENOTITLE]]

Date:3/18/2018

'''PCR of 3165 F3'''

Primers:3165 F1+3165 R2

Template:F1+F2

2XPCR Mix:Platinum

Extension time:30s

Annealing:55oC

'''Results:Cut out band and do the next step'''

[[File:IMG 3619.JPG|RTENOTITLE]]

'''Date:3/21/2018'''

'''Miniprep of 3165pMiniT2_3165_3131'''

3165pMiniT2:1-5 EcoRI

3165:6-10 BsrGI+NdeI

3131:11-15 BssHII+XhoI

Time:30min

'''Result:3165pMiniT2(1-5) have bands,3131(12-15)have bands.redo 3165 digestion.'''

[[File:IMG 3637.JPG|RTENOTITLE]]

Date:3/21/2018

'''Redo digestion of 3165 from PCR'''

Restriction enzymes:BamHI+BsrGI

Length:151bp

Time:15min

Buffer:Fastsdigest

'''Result:#2,#4 have good bands'''

[[File:IMG 3639.JPG|RTENOTITLE]]

'''Screening of miniprep_3165'''

2,4: BsrGI+SalI

2'',4'':BsrGI

Time:15Min

'''Result:seems good, Smear may caused by high concentration, around 300 ng/ul.'''

[[File:IMG 3642.JPG|RTENOTITLE]]

Date:2/26/2018

'''Test of 947_945_3166_3165'''

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| Name
| Enzymes
| Size (bp)
| Lane
|-
| 947
| BglII
| 646
| 1,2,3
|-
| <br/>
| BssHII
| 1856
| 4,5,6
|-
| 945
| BglII
| 628
| 11,12
|-
| <br/>
| BssHII
| 1838
| 13
|-
| 3165
| SalI+BsrGI
| 693
| 8
|-
| <br/>
| HindIII+BsrGI
| 441
| 10
|-
| 3166
| SalI+BsrGI
| 1234
| 9
|-
| <br/>
| XbaI
| 2049
| 7
|-
| <br/>
| BglII
| 3259
| to do
|}

[[File:IMG 36521.JPG|RTENOTITLE]]
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