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3165 pEF1-HTLV-1IFH-HA-VH-mIgG2b - Revision history
2026-10-18T07:54:58Z
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Crm at 15:54, 27 March 2018
2018-03-27T15:54:44Z
<p></p>
<p><b>New page</b></p><div>Test of 3165 miniprep<br />
<br />
Digest enzyme:BsrGI<br />
<br />
Time:30min<br />
<br />
Buffer:fastdigest<br />
<br />
Results:not good, got three bands<br />
<br />
[[File:Test of 3165.JPG|RTENOTITLE]]<br />
<br />
Test of 3165 miniprep digestion<br />
<br />
1:SalI+BsrGI NEB2<br />
<br />
2:SalI+KpnI Fastdigest<br />
<br />
3:BsrGI NEB2<br />
<br />
Time:30min<br />
<br />
[[File:Test3165.JPG|RTENOTITLE]]<br />
<br />
== Redo ==<br />
<br />
'''3165 PCR 1st'''<br />
<br />
F1:3028 as template<br />
<br />
Primer:3165F1+3165R1<br />
<br />
length440bp<br />
<br />
extenstion time:1:30<br />
<br />
F2:3129 as template<br />
<br />
Primer:3165F2+3165R2<br />
<br />
Buffer:Platinum<br />
<br />
length:165bp<br />
<br />
Extension time:15bp<br />
<br />
[[File:3165PCRF1+F2.JPG|RTENOTITLE]]<br />
<br />
Date:3/18/2018<br />
<br />
'''PCR of 3165 F3'''<br />
<br />
Primers:3165 F1+3165 R2<br />
<br />
Template:F1+F2<br />
<br />
2XPCR Mix:Platinum<br />
<br />
Extension time:30s<br />
<br />
Annealing:55oC<br />
<br />
'''Results:Cut out band and do the next step'''<br />
<br />
[[File:IMG 3619.JPG|RTENOTITLE]]<br />
<br />
'''Date:3/21/2018'''<br />
<br />
'''Miniprep of 3165pMiniT2_3165_3131'''<br />
<br />
3165pMiniT2:1-5 EcoRI<br />
<br />
3165:6-10 BsrGI+NdeI<br />
<br />
3131:11-15 BssHII+XhoI<br />
<br />
Time:30min<br />
<br />
'''Result:3165pMiniT2(1-5) have bands,3131(12-15)have bands.redo 3165 digestion.'''<br />
<br />
[[File:IMG 3637.JPG|RTENOTITLE]]<br />
<br />
Date:3/21/2018<br />
<br />
'''Redo digestion of 3165 from PCR'''<br />
<br />
Restriction enzymes:BamHI+BsrGI<br />
<br />
Length:151bp<br />
<br />
Time:15min<br />
<br />
Buffer:Fastsdigest<br />
<br />
'''Result:#2,#4 have good bands'''<br />
<br />
[[File:IMG 3639.JPG|RTENOTITLE]]<br />
<br />
'''Screening of miniprep_3165'''<br />
<br />
2,4: BsrGI+SalI<br />
<br />
2'',4'':BsrGI<br />
<br />
Time:15Min<br />
<br />
'''Result:seems good, Smear may caused by high concentration, around 300 ng/ul.'''<br />
<br />
[[File:IMG 3642.JPG|RTENOTITLE]]<br />
<br />
Date:2/26/2018<br />
<br />
'''Test of 947_945_3166_3165'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| Name<br />
| Enzymes<br />
| Size (bp)<br />
| Lane<br />
|-<br />
| 947<br />
| BglII<br />
| 646<br />
| 1,2,3<br />
|-<br />
| <br/><br />
| BssHII<br />
| 1856<br />
| 4,5,6<br />
|-<br />
| 945<br />
| BglII<br />
| 628<br />
| 11,12<br />
|-<br />
| <br/><br />
| BssHII<br />
| 1838<br />
| 13<br />
|-<br />
| 3165<br />
| SalI+BsrGI<br />
| 693<br />
| 8<br />
|-<br />
| <br/><br />
| HindIII+BsrGI<br />
| 441<br />
| 10<br />
|-<br />
| 3166<br />
| SalI+BsrGI<br />
| 1234<br />
| 9<br />
|-<br />
| <br/><br />
| XbaI<br />
| 2049<br />
| 7<br />
|-<br />
| <br/><br />
| BglII<br />
| 3259<br />
| to do<br />
|}<br />
<br />
[[File:IMG 36521.JPG|RTENOTITLE]]<br />
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