Paul.xu at 21:02, 18 October 2019

2019-10-18T21:02:57Z

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Cloning of pET42b-scIHF2(1203) Purpose: to clone pET42b-scIHF2 Date: 2012-5-23 to 2012-5-25 Notebook: Xiaojing Wang Page: 004252 Clone ID: 1203 Experimenters: Xiaojing Wang Results

Digest pET42b with Nco1 + Xho1--> vector digest 1078 pET28b-scIHF2 with Nco1 + Xho1--> insert 627bp

[[File:1202 1203 i v 5.23.2012.JPG|RTENOTITLE]]

Ligate and transform Select 6 clones to mini-prep 1203 plasmids are digested by Nco1 + Xho1

[[File:1203 digest 5.25.2012.JPG|RTENOTITLE]]

1203-1,-2,-3,-5 may be correct.
== 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]]

1203 pET42b-scIHF2 - Revision history

Paul.xu at 21:02, 18 October 2019