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2196-pT7A3E-PBCV-1-Lig - Revision history
2026-10-18T08:49:20Z
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Paul.xu at 20:28, 2 October 2017
2017-10-02T20:28:21Z
<p></p>
<p><b>New page</b></p><div>== '''Cloning of 2196-pT7A3E-PBCV-1-Lig''' ==<br />
<br />
'''Purpose:''' Cloning of 2196-pT7A3E-PBCV-1-Lig<br />
<br />
'''Date: '''2014-5-5---2014-5-8<br />
<br />
'''Notebook:0'''<br />
<br />
'''Page:'''<br />
<br />
'''Clone ID:2196'''<br />
<br />
'''Experimenters:''' yingying<br />
<br />
== '''Results''' ==<br />
<br />
=== cloning ===<br />
<br />
[[File:2196pcr 5-6-2014.JPG|RTENOTITLE]]<br />
<br />
gene synthesis<br />
<br />
PBCV-1-Lig-1 to PBCV-1-Lig-24<br />
<br />
[[File:2195 2196clone 5-6-2014.JPG|RTENOTITLE]]<br />
<br />
{| width="200" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| lane<br/><br />
| 3<br/><br />
| 4<br/><br />
|-<br />
| <br/><br />
| PCR<br/><br />
| 1892 pT7A3E<br/><br />
|-<br />
| <br/><br />
| colspan="2" | Nde1 + Xho1<br/><br />
|}<br />
<br />
<br />
<br />
=== Colony Screening ===<br />
<br />
Date: 2014-5-8<br />
<br />
Make miniprep and digest with Nde1 + Xho1<br />
<br />
[[File:2196digest 5-8-2014.JPG|RTENOTITLE]]<br />
<br />
&lt;br&gt;<br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
== '''Conclusion''' ==<br />
<br />
<br />
<br />
== '''Fix of 2196-pT7A3E-PBCV-1-Lig''' ==<br />
<br />
'''Purpose:''' fix of 2196-pT7A3E-PBCV-1-Lig<br />
<br />
'''Date: '''2014-5-12---2014-5-14<br />
<br />
'''Notebook:0'''<br />
<br />
'''Page:'''<br />
<br />
'''Clone ID:2196'''<br />
<br />
'''Experimenters:''' yingying<br />
<br />
== '''Results''' ==<br />
<br />
=== cloning ===<br />
<br />
[[File:2196pcr 5-12-2014.JPG|RTENOTITLE]]<br />
<br />
pcr1: tmplate 2196-1<br />
<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; PBCV-1-Lig-1+PBCV-1-Lig-10<br />
<br />
pcr2: tmplate 2196-2<br />
<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; PBCV-1-Lig-11+PBCV-1-Lig-22<br />
<br />
pcr3: tmplate 2196-1<br />
<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; PBCV-1-Lig-23+PBCV-1-Lig-23<br />
<br />
[[File:2196PCR 5-13-2014.JPG|RTENOTITLE]]<br />
<br />
PCR:template:pcr1+pcr2+pcr3<br />
<br />
&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; PBCV-1-Lig-1+PBCV-1-Lig-24<br />
<br />
[[File:1149+2196clone 5-13-2014.JPG|RTENOTITLE]]<br />
<br />
<br />
<br />
<br />
<br />
{| width="200" cellspacing="1" cellpadding="1" border="1"<br />
|-<br />
| lane<br/><br />
| 3<br/><br />
| 4<br/><br />
|-<br />
| <br/><br />
| PCR<br/><br />
| 1892 pT7A3E<br/><br />
|-<br />
| <br/><br />
| colspan="2" | Nde1 + Xho1<br/><br />
|}<br />
<br />
<br />
<br />
=== Colony Screening ===<br />
<br />
Date: 2014-5-15<br />
<br />
Make miniprep and digest with Nde1 + Xho1<br />
<br />
[[File:2196digest 5-15-2014.JPG|RTENOTITLE]]<br />
<br />
&lt;br&gt;<br />
<br />
== '''Result intepretation/Summary''' ==<br />
<br />
== '''Conclusion''' ==<br />
<br />
= Product info =<br />
<br />
[http://molbiol.edu.ru/eng/scripts/01_04.html http://molbiol.edu.ru/eng/scripts/01_04.html]<br />
<br />
1&nbsp;reaction require&nbsp; 8 pmol enzyme, 34k Da, 1 µl (25U), 1 pmole=34 ng, 272 ng--> 8 pmol.<br />
<br />
1&nbsp;µl -->25 U--8 pmole--> 272 ng<br />
[[Category:Cloning]]1,000 Units/25=40 ul. X272 ng =10&nbsp; ug /0.9 (90% purity)=12 ug.</div>
Paul.xu