Paul.xu at 18:17, 12 February 2013

2013-02-12T18:17:33Z

New page

== '''Cloning of 1546 PiggyBacHR-SMN1-5HA''' ==

'''Purpose:''' Cloning of 1546 PiggyBacHR-SMN1-5HA

'''Date: '''2013-1-28---2013-1-30

'''Notebook:0'''

'''Page:'''

'''Clone ID:1546'''

'''Experimenters:''' Shanyi

== '''Results''' ==

=== PCR ===

PCR from 293 genomic DNA digest with Nhe1 + BsaI clone into 1507 PiggyBacHR 

[[Image:1532 1561 1480 1546 1547pcr 1 25 2013.JPG]] 

[[Image:DIGEST FOR 1546.JPG]] 

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| band 1,2 
| band 3,4,5 
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| 1507 
| 1546PCR 
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| enzymes 
| colspan="2" |            Nhe1 + BsaI 
|}

 

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=== Colony Screening ===

Date: 2013-

Make miniprep and digest with Nhe1 + BsaI.

[[Image:1546DIGEST.JPG]]

[[Image:1546DIGEST WITH XbaI.JPG]]

NO ONE IS RIGHT 

 

== '''Cloning of 1546 PiggyBacHR-SMN1-5HA''' ==

'''Purpose:''' Cloning of 1546 PiggyBacHR-SMN1-5HA

'''Date: '''2013-1-28---2013-1-30

'''Notebook:0'''

'''Page:'''

'''Clone ID:1546'''

'''Experimenters:''' Shanyi

== '''Results''' ==

=== PCR ===

PCR from 293 genomic DNA 

[[Image:1546PCR 3rd TIME.JPG]] Method 1: digest 1/3 PCR products and 1507 PiggyBacHR with Nhe1 + BsaI(37oC 1hour, 50oC 1hour),then ligate 

Method2: digest and ligate together 10x (37 ̊C for 5min + 16 ̊C for 10min) + 50 ̊C for 5min + 80 ̊C for 5min 

=== Colony Screening ===

 [[Image:1546PCR SCREEN.JPG]]

The first 6 clone are from method2, the other 6 are from method 1. 

Make miniprep and digest with Nhe1 + BsaI. 

[[Image:1546digest.JPG]]

use plasmid 1234567as template

[[Image:1546pcr use plasmid as template.JPG]]

 

 

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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[[Category:Cloning]]

1546 PiggyBacHR-SMN1-5HA - Revision history

Paul.xu at 18:17, 12 February 2013