User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/12/20

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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
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