Difference between revisions of "User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/12/20"

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! Components !! No DNA Ctrl. (μl) !! GFP Ctrl. (μl) !! Target Protein (μl) with MV2 !! Target Protein (μl) with pT-CHis
 
! Components !! No DNA Ctrl. (μl) !! GFP Ctrl. (μl) !! Target Protein (μl) with MV2 !! Target Protein (μl) with pT-CHis
 
|-
 
|-
|format modifier (not displayed) | Hela Lysate ||43125
+
|format modifier (not displayed) | Hela Lysate ||12.5 || 12.5 || 12.5 || 12.5
 
|-
 
|-
|format  |Accessory Proteins ||format | 16143
+
|format  |Accessory Proteins ||format | 2.5 || 2.5 || 2.5 || 2.5
 
|-
 
|-
|format  | Reaction Mix ||format | 7978
+
|format  | Reaction Mix ||format | 5 || 5 || 5 || 5
 
|-
 
|-
|format  |pCFE-GFP DNA (0.5μg/μL) ||format | 4124
+
|format  |pCFE-GFP DNA (0.5μg/μL) ||format | --- || 2 || --- || ---
 
|-
 
|-
|format  | Cloned DNA (0.5 μg/μL) ||format | 2718
+
|format  | Cloned DNA (0.5 μg/μL) ||format | --- || --- || 5 || 3.3
 
|-
 
|-
|format | Nuclease free Water ||format|
+
|format | Nuclease free Water ||format| 5 || 3 || 0 || 1.7
 
|}
 
|}

Revision as of 17:14, 20 December 2012

Owwnotebook icon.pngPcTF Subcloning in E. coli <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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12/20/2012

1-Step Human Coupled in vitro protein expression

  • 1-Step Human In Vitro Protein Expression Kits enable the translation and post-transcriptional modification of full-length proteins from mRNA or plasmid templates with yields of up to 100µg/mL per reaction.

Kit Specifications

  • The PcTF gene with two different backbones were used for the protein expression.
    • pCFE1-CHis Expression vector
    • Mv2 vector , Part:BBa_J176122

Table of Components for the IVT reaction

Components No DNA Ctrl. (μl) GFP Ctrl. (μl) Target Protein (μl) with MV2 Target Protein (μl) with pT-CHis
Hela Lysate 12.5 12.5 12.5 12.5
Accessory Proteins 2.5 2.5 2.5 2.5
Reaction Mix 5 5 5 5
pCFE-GFP DNA (0.5μg/μL) --- 2 --- ---
Cloned DNA (0.5 μg/μL) --- --- 5 3.3
Nuclease free Water 5 3 0 1.7