Paul.xu at 19:26, 28 November 2012

2012-11-28T19:26:33Z

New page

== '''Cloning of 240-pET28b-T4-Ligase''' ==

'''Purpose:''' Cloning of 240-pET28b-T4-Ligase

'''Date: '''2012-10-11---2012-10-15

'''Notebook:0'''

'''Page:'''

'''Clone ID:240'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

1st PCR:Gene synthesis Frag 1: 1-14 Frag 2: 15-30 Frag 3: 31-44 

[[Image:240 PCR1+2+3 10-11-2012.JPG]][[Image:240 PCR 2 10-11-2012.JPG]] 

 

2nd PCR 

[[Image:240 PCR 10-12-2012.JPG]] 

 

[[Image:240+1097+1107+1352+1423 clone 10-12-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 281px; height: 66px;"
|-
| 
| band 1 
| band 2 
|-
| 
| PCR product 
| pET28b 
|-
| enzymes 
| colspan="2" |          Nde1+Xho1  
|}

 

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|-
| 
|}

=== Colony Screening ===

Date: 2012-10-15

Make miniprep and digest with Nde1+Xho1 

[[Image:240+1097+1107 digest detect 10-15-2012.JPG]] 

c 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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== '''Cloning of 240-pET28b-T4-Ligase''' ==

'''Purpose:''' Cloning of 240-pET28b-T4-Ligase

'''Date: '''2012-10-19---2012-10-21

'''Notebook:0'''

'''Page:'''

'''Clone ID:240'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

2nd PCR:2012-10-19

[[Image:240+1095+1248 PCR 10-19-2012.JPG]]

 

2012-10-23

[[Image:240+1095 clone 10-23-2012.JPG]] 

{| width="200" cellspacing="1" cellpadding="1" border="1"
|-
| 
| lane1 
| 
|-
| 
| PCR product 
| 
|-
| enzymes 
| Nde1+Xho1 
| 
|}

 

2012-10-24

[[Image:240+1095+1420 clone 10-24-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 281px; height: 66px;"
|-
| 
| 
| lane1 
|-
| 
| 
| 1026,pET28b-LambdaExo 
|-
| enzymes 
| colspan="2" |          Nde1+Xho1  
|}

 

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

=== Colony Screening ===

Date: 2012-10-21

Make miniprep and digest with Nde1+Xho1 

[[Image:240+1095 digest detect 10-26-2012.JPG]]

clone 5 need to sequence

 

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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== '''Cloning of 240-pET28b-T4-Ligase''' ==

'''Purpose:''' Cloning of 240-pET28b-T4-Ligase

'''Date: '''2012-11-20---2012-10-28

'''Notebook:0'''

'''Page:'''

'''Clone ID:240'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

2nd PCR:2012-10-19

[[Image:240+1095+1248 PCR 10-19-2012.JPG]]

 

2012-11-20 

[[Image:240+1096+1463_clone_11-20-2012.JPG]] 

{| width="410" height="84" cellspacing="1" cellpadding="1" border="1"
|-
| 
| lane 1 
| lane2 
|-
| 
| PCR product 
| 1027,pET28b-Lambda 
|-
| enzymes 
| colspan="2" |          Nde1+Xho1  
|}

 

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

=== Colony Screening ===

Date: 2012-11-27

Make miniprep and digest with Nde1+Xho1 

[[Image:240_digest_detect_11-28-2012.JPG]] 

PCR screening:T4 1-1+T7tem

[[Image:240_PCR_screen_11-27-2012.JPG]]

 

 

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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[[Category:Cloning]]

240-pET28b-T4-Ligase - Revision history

Paul.xu at 19:26, 28 November 2012