Editing 1144 pT7A3-MBP-(3C)-Bst-Exo- (section)

== '''Results'''  ==

=== cloning  ===

Date:2013-3-<br> 

Gene synthesis 3 PCR fragments 

[[Image:1144-1 -2 -3PCR.JPG]] 

<br>&nbsp;then use Fragment1,2,3 as template 

Bst-Exo-1-1+Bst-Exo-3-18 as primers 

[[Image:1144PCR.JPG]]<br> 

digest&nbsp;: 

[[Image:Digest for 1144pec.JPG]]<br> 

{| cellspacing="1" cellpadding="1" border="1" style="width: 298px; height: 82px;"
|-
| <br> 
| band 1<br> 
| band 3<br>
|-
| <br> 
| &nbsp; PCR product<br> 
| 1454<br>
|-
| enzymes<br> 
| colspan="2" | &nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Nde1+ Xho1<br>
|}

<br> 

{| cellspacing="1" cellpadding="1" border="1"
|-
| <br>
|}

=== Colony Screening  ===

Date: 2013 

DIGEST WITH NdeI , XhoI and BamHI (should get there bands=&gt;350bp, 1500bp,2500bp) 

[[Image:1144pEC-Bst-ExoDIGEST.JPG]]<br> 

SEND 1144-1 TO SEQUENCE, missing 3nt 

send 1144-2 to sequence, missing 1G(SEQUENCE FROM T7 TERM AND M13R)<br> 

<br> 

=== cloning  ===

Date:2013-3-<br> 

Fragment 1, use Bst-Exo-1-1 and Bst-Exo-3-12 

Fragment 2,use Bst-Exo-3-13 and Bst-Exo-3-18<br> 

[[Image:1144fix 1705pcr 2013 04 10.JPG]]<br>&nbsp;then use Fragment 1and2 as template 

Bst-Exo-1-1+Bst-Exo-3-18 as primers 

[[Image:1144fix 1048fix 2013 04 11.JPG]]<br> 

digest&nbsp;: 

[[Image:Digest for 1144 04 11 2013.JPG]]<br> 

<br> 

=== Colony Screening  ===

Date: 2013 

DIGEST WITH NdeI , XhoI and BamHI (should get there bands=&gt;350bp, 1500bp,2500bp) 

[[Image:1313 1314 1144FIX DIGEST.JPG]][[Image:1144FIX DIGEST.JPG]]<br> 

SEND 1144FIX-5 TO SEQUENCE, MISS ONE T and ONE A,NEED TO FIX THEM<br> 

=== cloning  ===

Date:2013-3-<br> 

Fragment 1, use Bst-Exo-1-1 and Bst-Exo-2-8 

Fragment 2,use Bst-Exo-2-9 and Bst-Exo-3-18<br> 

[[Image:1144fixthe missing AT using1144fix-5as template.JPG]]<br>&nbsp;cut out the right bands, then use Fragment 1and2 as template 

Bst-Exo-1-1+Bst-Exo-3-18 as primers 

[[Image:1144fixthe missing AT.JPG]]<br> 

digest&nbsp;: 

[[Image:Digest for1144fixthe missing AT.JPG]]<br> 

<br> 

=== Colony Screening  ===

Date: 2013 

DIGEST WITH NdeI , XhoI and BamHI (should get there bands=&gt;350bp, 1500bp,2500bp) 

[[Image:1144fixthe missing ATDIGEST.JPG]]<br> 

[[Image:562fix_622_1144fix3_1740digest.JPG]]

<br>