Revision as of 20:06, 20 December 2012

PcTF Subcloning in E. coli

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.

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	43125
Accessory Proteins	16143
Reaction Mix	7978
pCFE-GFP DNA (0.5μg/μL)	4124
Cloned DNA (0.5 μg/μL)	2718
Nuclease free Water

@@ Line 18: / Line 18: @@
 '''Table of Components for the IVT reaction'''
+{| border="1"
+! 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  |Accessory Proteins ||format | 16143
+|-
+|format  | Reaction Mix ||format | 7978
+|-
+|format  |pCFE-GFP DNA (0.5μg/μL) ||format | 4124
+|-
+|format  | Cloned DNA (0.5 μg/μL) ||format | 2718
+|-
+|format | Nuclease free Water ||format|
+|}