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Yijia LI 7 23 2018 - Revision history
2026-10-18T06:28:23Z
Revision history for this page on the wiki
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Crm at 17:22, 1 August 2018
2018-08-01T17:22:17Z
<p></p>
<p><b>New page</b></p><div>== Date:7/23/2018 ==<br />
<br />
'''Miniprep of 931_3206_129'''<br />
<br />
[[File:IMG 1177.JPG|RTENOTITLE]]<br />
<br />
== Date:7/24/2018 ==<br />
<br />
'''931 subclone'''<br />
<br />
[[File:IMG 1180.JPG]]<br />
<br />
== Date:7/25/2918 ==<br />
<br />
'''3218 miniprep digestion'''<br />
<br />
[[File:IMG 1198.JPG|RTENOTITLE]]<br />
<br />
== Date:7/26/2918 ==<br />
<br />
'''3206 PCR screening 1'''<br />
<br />
'''PCR use 2x homemade Taq mix'''<br />
<br />
[[File:3206pcrscreening.JPG|RTENOTITLE]]<br />
<br />
'''3206 PCR screening 2'''<br />
<br />
'''PCR use 2x homemade Taq mix'''<br />
<br />
[[File:3206pcrscreening2.JPG|RTENOTITLE]]<br />
<br />
'''0.1Taq QC test'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #Lane<br />
| 1<br />
| 2<br />
| 3<br />
| 4<br />
| 5<br />
| 6<br />
| 7<br />
| 8<br />
| 9<br />
| 10<br />
| 11<br />
| 12<br />
|-<br />
| Name<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 0.1Taq<br />
| 2X BestTaq Mastermix<br />
| Excellgen<br />
|-<br />
| Concentration<br />
| 880ng/ul<br />
| 220ng/ul<br />
| 110ng/ul<br />
| 55ng/ul<br />
| 27.5ng/ul<br />
| 13.75<br />
| 6.875<br />
| 3.43<br />
| 1.71<br />
| 0.85<br />
| + control<br />
| +control<br />
|}<br />
<br />
[[File:IMG 1209.JPG|RTENOTITLE]]<br />
<br />
'''miniprep digestion 3225_999_931'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| Vector<br />
| Enyzmes<br />
| Size<br />
| note<br />
|-<br />
| 3225<br />
| 1-4; 5 is + control<br />
| 1851 pT7A3-GST<br />
| Nhe1+Xho1<br />
| 383bp<br />
| <br/><br />
|-<br />
| 999<br />
| 6-9;10 is +control<br />
| 1873<br />
| Nde1+Xho1<br />
| 1182bp<br />
| <br/><br />
|-<br />
| 931<br />
| 11-14;15 is + control<br />
| 931-1<br />
| HindIII+Xho1<br />
| <br/><br />
| failed<br />
|}<br />
<br />
[[File:IMG 1211.JPG|RTENOTITLE]]<br />
<br />
'''Miniprep digestion of 941_3244_998'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| Vector<br />
| Method<br />
| style="width: 106px;" | Enzymes<br />
| style="width: 34px;" | Size<br />
| style="width: 34px;" | Note<br />
|-<br />
| 941<br />
| 1-4; 5&nbsp; is + control<br />
| 947<br />
| Subclone<br />
| style="width: 106px;" | Nde1+Xho1<br />
| style="width: 34px;" | 456bp<br />
| style="width: 34px;" | <br/><br />
|-<br />
| 3244<br />
| 6-9<br />
| 3242<br />
| PCR cloning<br />
| style="width: 106px;" | Nde1+HindIII<br />
| style="width: 34px;" | 739bp<br />
| style="width: 34px;" | vector,wrong<br />
|-<br />
| 998<br />
| 10-13<br />
| 1708<br />
| Gibson Assembly<br />
| style="width: 106px;" | HindIII+Xho1<br />
| style="width: 34px;" | <br/><br />
| style="width: 34px;" | <br/><br />
|}<br />
<br />
[[File:IMG 1213.JPG|RTENOTITLE]]<br />
<br />
== Date:7/27/2018 ==<br />
<br />
'''2nd miniprep digestion_3225_999_998'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| Enzymes<br />
| Vector<br />
| Size<br />
| Note<br />
|-<br />
| 3225<br />
| 1-4<br />
| Nco1+Xho1<br />
| 1851<br />
| three fragment.5288bp;738bp;361bp<br />
| #3,#4<br />
|-<br />
| 999<br />
| 5-8<br />
| HindIII+Xho1<br />
| 1873<br />
| 1115bp<br />
| <br/><br />
|-<br />
| 998<br />
| 9-12<br />
| HindIII+Kpn1<br />
| 1708<br />
| should have four fragments<br />
| Failed<br />
|}<br />
<br />
[[File:IMG 1226.JPG|RTENOTITLE]]<br />
<br />
'''miniprep digestion_3249_3206_931-1'''<br />
<br />
{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"<br />
|-<br />
| #<br />
| Lane<br />
| Vector<br />
| Enyzmes<br />
| Size<br />
| note<br />
|-<br />
| 3249<br />
| 1-4; 5 is + control;6 is vector - control<br />
| 1834<br />
| Sph1+Xho1<br />
| 1156bp<br />
| <br/><br />
|-<br />
| 3206<br />
| 7-10<br />
| 3165<br />
| BamH1+HindIII<br />
| <br/><br />
| Failed<br />
|-<br />
| 931-1<br />
| 11-14; 15 is +control, 16 is - control<br />
| 927<br />
| Nde1+HindIII<br />
| ~989bp<br />
| lane 11,lane 12<br />
|}<br />
<br />
[[File:IMG 1224.JPG|RTENOTITLE]]<br />
[[Category:My ELN]]</div>
Crm