3165 pEF1-HTLV-1IFH-HA-VH-mIgG2b
Test of 3165 miniprep
Digest enzyme:BsrGI
Time:30min
Buffer:fastdigest
Results:not good, got three bands
Test of 3165 miniprep digestion
1:SalI+BsrGI NEB2
2:SalI+KpnI Fastdigest
3:BsrGI NEB2
Time:30min
Redo
[edit]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
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
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.
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
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.
Date:2/26/2018
Test of 947_945_3166_3165
| Name | Enzymes | Size (bp) | Lane |
| 947 | BglII | 646 | 1,2,3 |
| BssHII | 1856 | 4,5,6 | |
| 945 | BglII | 628 | 11,12 |
| BssHII | 1838 | 13 | |
| 3165 | SalI+BsrGI | 693 | 8 |
| HindIII+BsrGI | 441 | 10 | |
| 3166 | SalI+BsrGI | 1234 | 9 |
| XbaI | 2049 | 7 | |
| BglII | 3259 | to do |
