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942 pET28b-pMBP-VVN - Revision history
2026-10-18T11:23:30Z
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Crm at 20:14, 19 April 2018
2018-04-19T20:14:59Z
<p></p>
<p><b>New page</b></p><div>[[File:938 939 942 3122 Nde1 Nco1 Xho1.JPG|RTENOTITLE]]<br />
<br />
[[File:938 939 942 3122 Xho1 HindIII.JPG|RTENOTITLE]]<br />
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12/27/2017<br />
<br />
938 digestion with Nde1-Xho1, Fastdigest Buffer<br />
<br />
[[File:938 939 942 3122 931 1855 digestion.JPG|RTENOTITLE]]<br />
<br />
{| 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 />
<br />
[[File:938 939 942 3122 931 1855 digestion Cutsmart.JPG|RTENOTITLE]]<br />
<br />
== Date:4/5/2018 ==<br />
<br />
'''subclone of 942'''<br />
<br />
'''Size:716bp'''<br />
<br />
'''Vector:947 pT7A3'''<br />
<br />
'''Insert:942'''<br />
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[[File:Subclone942.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep digestion of 984_942_824_3166'''<br />
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{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| Size<br />
| Enzymes<br />
| Note<br />
|-<br />
| 984<br />
| 1-4<br />
| <br/><br />
| Xba1+NdeI<br />
| Subclone<br />
|-<br />
| 942<br />
| 5-8<br />
| 716bp<br />
| <div><div>Nco1-Xho1</div></div><br />
| <br/><br />
|-<br />
| 824<br />
| 9-12<br />
| 969<br/><br />
| Nde1 + Xho1<br/><br />
| <br/><br />
|-<br />
| 3166<br />
| 13-16<br />
| 1234bp<br />
| Sal1+BsrG1<br/><br />
| PCR<br />
|}<br />
<br />
<br />
<div>&nbsp;[[File:IMG 0659.JPG|RTENOTITLE]]</div><br />
'''test for selecting correct subclone of 986_944_945_942_824'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width: 500px;"<br />
|-<br />
| #<br />
| Lane<br />
| control<br />
| Size<br />
| style="width: 65px;" | Enzymes<br />
| style="width: 195px;" | Note<br />
|-<br />
| 986<br />
| 1-2<br />
| Lane 3 1708 vector<br />
| <br/><br />
| style="width: 65px;" | HindIII+XhoI<br/><br />
| style="width: 195px;" | undigested vector<br />
|-<br />
| 944<br />
| 4-6<br />
| Lane 7&nbsp; 944pMIniT2<br />
| 2334bp<br />
| style="width: 65px;" | NdeI+XhoI<br/><br />
| style="width: 195px;" | from PCR, '''No.4 might be right one'''<br />
|-<br />
| 945<br />
| 8<br />
| Lane 9&nbsp; 945pMiniT2<br />
| 1326bp<br />
| style="width: 65px;" | Xba1-Nco1<br/><br />
| style="width: 195px;" | '''good'''<br />
|-<br />
| 942<br />
| 10-12<br />
| Lane 13 942pJet1.2<br />
| 716bp<br />
| style="width: 65px;" | Nco1-Xho1<br/><br />
| style="width: 195px;" | '''good'''<br />
|-<br />
| 824<br />
| 14-15<br />
| Lane 16 832pMiniT2<br />
| 969bp<br />
| style="width: 65px;" | Nde1 + Xho1<br/><br />
| style="width: 195px;" | <br/><br />
|}<br />
<br />
[[File:Redo test for miniprep.JPG|RTENOTITLE]]<br />
[[Category:Cloning]]</div>
Crm