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938 pET30b-PNGase F - Revision history
2026-10-18T06:29:47Z
Revision history for this page on the wiki
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Crm at 20:23, 19 April 2018
2018-04-19T20:23:24Z
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<p><b>New page</b></p><div>[[File:938 939 942 3122 Nde1 Nco1 Xho1.JPG|RTENOTITLE]]<br />
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[[File:938 939 942 3122 Xho1 HindIII.JPG|RTENOTITLE]]<br />
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12/27/2017<br />
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938 digestion with Nde1-Xho1, Fastdigest Buffer<br />
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[[File:938 939 942 3122 931 1855 digestion.JPG|RTENOTITLE]]<br />
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{| border="1" cellspacing="1" cellpadding="1" style="width: 681px;"<br />
|-<br />
| 1<br />
| 2<br />
| 3<br />
| 4<br />
| 5<br />
| 6<br />
| 7<br />
| style="width: 54px;" | 8,9<br />
| style="width: 55px;" | 10,11<br />
| style="width: 71px;" | 13<br />
|-<br />
| 938&nbsp;PNGase_F<br />
| <span style="color: rgb(0, 0, 0); font-family: Arial, Verdana, sans-serif; font-size: 12px; white-space: nowrap;">939 Tte-ExoIII</span><br/><br />
| 942 VVN<br/><br />
| 3122 CRM197<br/><br />
| 931, 1, PAK16D ligase&nbsp;<br/><br />
| 931, 2, PAK16D ligase<br/><br />
| 1855&nbsp; yPolyA-Pol<br/><br />
| style="width: 54px;" | 1540<br />
| style="width: 55px;" | 2046<br />
| style="width: 71px;" | 2046<br/><br />
|-<br />
| Nde1-Xho1<br />
| Nde1-Xho1<br/><br />
| <span style="color:#FF0000;">Nco1-</span>Xho1<br/><br />
| Nde1-Xho1<br/><br />
| Nde1-<span style="color:#FF0000;">HindIII</span><br/><br />
| <span style="color:#FF0000;">HindIII</span>-Xho1<br />
| Nde1-Xho1<br />
| style="width: 54px;" | Nde1-Xho1<br />
| style="width: 55px;" | Nde1-Xho1<br />
| style="width: 71px;" | <span style="color:#FF0000;">Nco1</span>-Xho1<br />
|-<br />
| pJet1.2<br />
| pJet1.2<br/><br />
| pJet1.2<br/><br />
| pJet1.2<br/><br />
| pJet1.2<br/><br />
| pJet1.2<br/><br />
| pEC2-<br/><br />
| style="width: 54px;" | pET30b<br />
| style="width: 55px;" | pET28b<br />
| style="width: 71px;" | pET28b<br/><br />
|-<br />
| 1145<br/><br />
| 780<br/><br />
| 716<br/><br />
| 1798<br/><br />
| 1174<br/><br />
| 986<br/><br />
| 1713<br />
| style="width: 54px;" | there is no Xho1 site<br />
| style="width: 55px;" | 1377<br />
| style="width: 71px;" | ~1377<br />
|-<br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
| <br/><br />
| style="width: 54px;" | <br/><br />
| style="width: 55px;" | <br/><br />
| style="width: 71px;" | <br/><br />
|}<br />
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[[File:938 939 942 3122 931 1855 digestion Cutsmart.JPG|RTENOTITLE]]<br />
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== ==<br />
<br />
'''subclone of 938'''<br />
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Size:1145bp<br />
<br />
V:1987 pT7A3<br />
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[[File:IMG 3672.JPG|RTENOTITLE]]<br />
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== Date:3/30/2018 ==<br />
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'''Miniprep digestion of 3122_938_939'''<br />
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3122:lane 1-4 lane 5&nbsp;:+ size:1743 bp<br />
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938: lane 6-9 lane 10:+ size:1145 bp<br />
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939: lane 11-14 lane 15:+ size:780 bp<br />
<br />
Positive control are insert-pjet1.2<br />
<br />
The small band size of vector 1727:193 bp<br />
<br />
[[File:IMG 0535.JPG|RTENOTITLE]]<br />
<br />
'''redo Miniprep digestion of 3122_938_939'''<br />
<br />
1:3122&nbsp; size:1743bp<br />
<br />
2:3122pjet1.2 sequenced +<br />
<br />
3:938&nbsp; size:1145 bp<br />
<br />
4:938pjet1.2 sequenced +<br />
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5,6,7:939&nbsp; size:780 bp<br />
<br />
8:939pjet1.2 sequenced +<br />
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Enzymes:NdeI+XhoI<br />
<br />
[[File:IMG 0536.JPG|RTENOTITLE]]<br />
<br />
'''938 miniprep test'''<br />
<br />
1:938-1<br />
<br />
2:938pJet1.2<br />
<br />
Enzymes:EcoRV+XhoI<br />
<br />
Size:552bp<br />
<br />
3:938-1<br />
<br />
4:938pJet1.2<br />
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Enzymes:NdeI+XhoI<br />
<br />
Size:1071bp<br />
<br />
[[File:IMG 0619.JPG|RTENOTITLE]]<br />
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== Date:4/5/2018 ==<br />
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'''938 subclone'''<br />
<br />
'''Size:1145bp'''<br />
<br />
'''Insert:938pJet1.2'''<br />
<br />
'''Vector:1727 pT7A30'''<br />
<br />
'''Enzymes:Nde1+Xho1'''<br />
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[[File:938subclone.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep digestion of 938_939_984'''<br />
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{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| style="width: 34px;" | Enzymes<br />
| style="width: 43px;" | Size<br />
| Note<br />
|-<br />
| 938<br />
| 1-4;5 is + control(sequenced 938pjet1.2)<br />
| style="width: 34px;" | Nde1+Xho1<br />
| style="width: 43px;" | 1145bp<br />
| '''good'''<br />
|-<br />
| 939<br />
| 6-9&nbsp;; 10 is + control(sequenced 939pJet1.2)<br />
| style="width: 34px;" | Nde1+Xho1<br />
| style="width: 43px;" | 780bp<br />
| '''good'''<br />
|-<br />
| 984<br />
| 11-14; 15 is + control(sequenced 982)<br />
| style="width: 34px;" | Xba1+Nhe1<br />
| style="width: 43px;" | 957bp<br />
| need to redo<br />
|}<br />
<br />
[[File:Miniprepdigestion 938 939 984.JPG|RTENOTITLE]]<br />
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[[Category:Cloning]]</div>
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