Paul.xu at 14:02, 10 May 2012

2012-05-10T14:02:52Z

New page

== '''Cloning of 1017-pDNR-Dual-ATR-C-His''' ==

'''Purpose:''' Cloning of 1005-pBad24-Tat-sfGFP by PCR

'''Date: '''2012-3-21--2012-3-26

'''Notebook:0'''

'''Page:'''

'''Clone ID:1017'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

Date:2012-3-21

[[Image:1011+1017 PCR 3-21-2012.JPG]]

Template:1016 pDNR-Dual-ATR 

ATR-AgeI-F GCAATAAACCGGTTGATGGAAG

ATR-BssHII-R BamH1 BssHII (GGTTGGACTCCATATATGcatcatcaccatcaccatTAGGGATCCGCGCGCagctaca) tgtagctGCGCGCGGATCCCTAatggtgatggtgatgatgCATATATGGAGTCCAACC 

[[Image:1011+1017+1021clone 3-22-2012.JPG]]

{| width="200" cellspacing="1" cellpadding="1" border="1"
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| band 3 
| band 4 
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| PCR prodduct 
| pDNR-Dual-ATR 
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| enzymes 
| colspan="2" |       AgeI+BssHII 
|}

 

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=== Colony Screening ===

Date: 2012-3-26

Make miniprep and digest with AgeI+BssHII 

[[Image:1011+1017+1021 digest detect 3-26-2012.JPG]]

6,7,8,9,10 no insert ,need to clone again

 '''Result intepretation/Summary'''

== ==

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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== '''Cloning of 1017-pDNR-Dual-ATR-C-His''' ==

'''Purpose:''' Cloning of 1005-pBad24-Tat-sfGFP by PCR

'''Date: '''2012-3-27--2012-3-30

'''Notebook:'''

'''Page:003783'''

'''Clone ID:1017'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

Date:2012-3-21

[[Image:1011+1017 PCR 3-21-2012.JPG]]

Template:1016 pDNR-Dual-ATR 

ATR-AgeI-F GCAATAAACCGGTTGATGGAAG

ATR-BssHII-R BamH1 BssHII (GGTTGGACTCCATATATGcatcatcaccatcaccatTAGGGATCCGCGCGCagctaca) tgtagctGCGCGCGGATCCCTAatggtgatggtgatgatgCATATATGGAGTCCAACC 

digest: [[Image:1009+1017+1023+1034 clone 3-27-2012.JPG]] 

AgeI at 37c for 1 hour,then BssHII at 50c for 1 hour 

{| width="200" cellspacing="1" cellpadding="1" border="1"
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| band 3 
| band 4 
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| PCR prodduct 
| pDNR-Dual-ATR 
|-
| enzymes 
| colspan="2" |       AgeI+BssHII 
|}

 

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=== Colony Screening ===

Date: 2012-3-29

Make miniprep and digest with Xba1 

no band

 

 '''Result intepretation/Summary'''

== ==

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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== '''Cloning of 1017-pDNR-Dual-ATR-C-His''' ==

'''Purpose:''' Cloning of 1017-pDNR-Dual-ATR-C-His

'''Date: '''2012-4-13---2012-4-16

'''Notebook:0'''

'''Page:'''

'''Clone ID:1017'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

[[Image:1017+1095 PCR 4-13-2012.JPG]] 

template:1016 pDNR-Dual-ATR 

PCR using these primers

 ATR-AgeI-F GCAATAAACCGGTTGATGGAAG ATR-AvrII-R BamH1 AvrII (GGTTGGACTCCATATATGcatcatcaccatcaccatTAGGGATCCTAGGagctacaa) ttgtagctCCTAGGATCCCTAatggtgatggtgatgatgCATATATGGAGTCCAACC 

[[Image:1017 clone 4-20-2012.JPG]] 

'''vector can not de digested''' 

 

{| cellspacing="1" cellpadding="1" border="1" style="width: 417px; height: 82px;"
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| band 1 
| band 2 
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| PCR product 
| 1016,pDNR-Dual-ATR 
|-
| enzymes 
| colspan="2" |           AgeI+ AvrII 
|}

 

need to digest with Sal1 + age1 and Sal1 + BglII 

=== Colony Screening ===

 

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

 

== '''Conclusion''' ==

 

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== '''Cloning of 1017-pDNR-Dual-ATR-C-His''' ==

'''Purpose:''' Cloning of 1005-pBad24-Tat-sfGFP by PCR

'''Date: '''2012-5-9--2012-

'''Notebook:0'''

'''Page:'''

'''Clone ID:1017'''

'''Experimenters:''' yingying

== '''Results''' ==

=== cloning ===

Date:2012-5-9

[[Image:1017_vector_digest_5-9-2012.JPG]] 

{| cellspacing="1" cellpadding="1" border="1" style="width: 421px; height: 66px;"
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| band 1 
| band 2 
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| 1016,pDNR-Dual-ATR 
| 1016,pDNR-Dual-ATR 
|-
| rowspan="2" | enzymes 
| rowspan="2" |      Not1+Bgl11 
| rowspan="2" | Sal1+Bgl11
|-
|}

 

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=== Colony Screening ===

Date: 

 '''Result intepretation/Summary'''

== ==

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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&lt;br&gt;

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[[Category:Cloning]]

1017-pDNR-Dual-ATR-C-His - Revision history

Paul.xu at 14:02, 10 May 2012