Paul.xu at 18:29, 21 May 2012

2012-05-21T18:29:29Z

New page

May 2, 2012

PCR using template from gene synthesis

[[Image:1129 PCR after gene synthesis 5 2 2012.jpg]]

digest with Kpn1 + Nde1, clone into Kpn1+ Nde1 digested 360 pDB.His.MBP 

Select 6 clones to mini-prep 1129 plasmids are digested by Nde1 + Kpn1 

[[Image:1129 digested 5.4.2012.JPG]]

Select 1129-5 to sequence.

==

== ==

== '''Cloning of 1129 pDB.His.PfuMBP''' ==

'''Purpose:''' Cloning of 1129 pDB.His.PfuMBP

'''Date: '''2012-5-17---

'''Notebook:0'''

'''Page:'''

'''Clone ID:1129'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

 

[[Image:1128+1129+1073 clone 5-17-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 281px; height: 66px;"
|-
| 
| band 1 
| band 2 
|-
| 
| PCR product 
| 360 pDB.His.MBP 
|-
| enzymes 
| colspan="2" |             Kpn1 + Nde1 
|}

 

 

2012-5-18 

[[Image:1128+1129 clone 5-18-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 346px; height: 96px;"
|-
| 
| band 3 
| band 4 
|-
| 
|
PCR product 

| 360 pDB.His.MBP 
|-
| enzymes 
| colspan="2" |           Kpn1 + Nde1 
|}

 

{| cellspacing="1" cellpadding="1" border="1"
|-
| 
|}

=== Colony Screening ===

Date: 2012-5-21

Make miniprep and digest with Kpn1 + Nde1

[[Image:1128+1129_digest_detect_5-21-2012.JPG]] 

clone 1,2,3 need to sequence 

== '''Result intepretation/Summary''' ==

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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[[Category:Cloning]]

1129 pDB.His.PfuMBP - Revision history

Paul.xu at 18:29, 21 May 2012