Editing 736 pEC2-mNco1

== '''Cloning of 736 pEC2-mNco1'''  ==

'''Purpose:''' Cloning of736 pEC2-mNco1'''==''' 

'''Date: '''2014-9---- 

'''Notebook:0''' 

'''Page:''' 

'''Clone ID:736''' 

'''Experimenters:'''&nbsp; shanyi 

== '''Results'''  ==

=== CLONING  ===

gene synthesis 

[[Image:736 2351pcr.JPG]] 

jion two fragments by PCR 

[[Image:736 2351pcr-C.JPG]] 

DIGEST WITH NdeI+XhoI 

[[Image:DIGEST FOR736 2351.JPG]]<br> 

=== Colony Screening  ===

Date: 2014_8<br> 

Make miniprep, use NdeI + XhoI to find positive clone<br> 

[[Image:736 2351DIGEST.JPG]]<br> 

<br> 

=== fix 736-5 pEC2-mNco1  ===

pcr <br> 

[[Image:736fix 1916fix2 2323pcr.JPG]][[Image:736fix 1916fix2 2323pcr2.JPG]] 

jOIN two fragment by PCR 

[[Image:736fix 1916fix2 1917 2240.JPG]]<br> digest with Nde1 + XhoI<br> 

[[Image:Digest for2354 2369 736fix2 1916fix3 2240.JPG]][[Image:Digest for736fix2 1916fix3 2240.JPG]]<br> 

=== Colony Screening  ===

Date: 2014_8<br> 

Make miniprep, use&nbsp; Nde1 + Xho1 to find positive clone<br> 

[[Image:736FIX2 1916FIX3 2240 2354 2369 2375.JPG]]<br> 

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

*<br>

== '''Conclusion'''  ==

=== fix 736-5 pEC2-mNco1  ===

pcr&nbsp; with Tth-muts<br> 

[[Image:2401-1 2198 2229-1 736fix3-1pcr.JPG]][[Image:2229-2 736fix3-2pcr.JPG]]<br> 

jOIN two fragment by PCR 

[[Image:2229 736fix3pcr.JPG]]<br> digest with Nde1 + XhoI<br> 

[[Image:Digest for 736fix3 2198 2229 2416.JPG]]<br> 

=== Colony Screening  ===

Date: 2015_1<br> 

Make miniprep, use&nbsp; Nde1 + PST1 to find positive clone<br> [[Image:736fix3 2198 2229digest.JPG]]<br> 

<br> 

[[Category:Cloning]]