Paul.xu at 21:03, 18 October 2019

2019-10-18T21:03:23Z

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== '''Cloning of 1074 pET28b-IHF''' ==

'''Purpose:''' Cloning of 1074 pET28b-IHF

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

'''Notebook:0'''

'''Page:'''

'''Clone ID:1074'''

'''Experimenters:''' yingying

== '''Results''' ==

=== subcloning ===

[[File:1074 clone 4-13-2012.JPG|RTENOTITLE]]

{| cellspacing="1" cellpadding="1" border="1" style="width: 298px; height: 80px;"
|-
| 
| band 1 
| band 2 
|-
| 
| 405 pMCSG13-IHF 
| pET28b 
|-
| enzymes 
| colspan="2" | Nco1 + Xho1 
|}

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

=== Colony Screening ===

Date: 2012-4-16

Make miniprep and digest with Nco1 + Xho1.

[[File:1074 digest detect 4-16-2012.JPG|RTENOTITLE]]

 

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

*&lt;br&gt;

== '''Conclusion''' ==

&lt;br&gt;

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|}

&lt;br&gt;

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== Protein Expression ==

induction, 37 oC 5 hours

{| border="1" cellspacing="1" cellpadding="1" style="width:500px;"
|-
| Lane
| ID
| Name
| Pi, MW
| Induction
| Sample
| Expression
|-
| 1
| 1708
| pET28b-scIHF2 
| 
| 
| S
| 
|-
| 2
| 1203
| pET42b-scIHF2 
| 
| 
| S
| 
|-
| 3
| 1074
| pET28b-IHFab 
| 
| 
| S
| 
|-
| 4
| 1186
| pDB.His.MBP-Xis 
| 
| 
| S
| 
|-
| 5
| 406 
| pET28b-Int 
| 
| 
| S
| 
|-
| 6
| 1185
| pDB.His.MBP-Int 
| 
| 
| 
| 
|-
| 7
| 1708 
| pET28b-scIHF2 
| 9.60 / 22.8 
| 
| P
| Y
|-
| 8
| 1203 
| pET42b-scIHF2 
| 8.38 / 54.2 
| 
| P
| Y
|-
| 9
| 1074 
| pET28b-IHFab 
| 9.20 / 25.3 
| 
| P 
| N
|-
| 10
| 1186 
| pDB.His.MBP-Xis 
| 6.62 / 52.1 
| 
| P 
| Y
|-
| 11
| 406 
| pET28b-Int 
| 9.53 / 42.5 
| 
| P 
| N
|-
| 12
| 1185 
| pDB.His.MBP-Int 
| 7.33 / 83.7 
| 
| P 
| Y
|-
| M
| 
| PageRuler Plus
| rowspan="1" colspan="4" | 250, 130, 00, 70, 55, 35, 25, 15, 10
|}

[[File:IHF XIS INT Expression screening.jpg]]
[[Category:Cloning]]

1074 pET28b-IHF - Revision history

Paul.xu at 21:03, 18 October 2019