ExiProgen™ ProXpress PCR Template Kit


ExiProgen™ ProXpress PCR Template Kit is the most optimized product for preparing template DNA for protein synthesis on the ExiProgen™.



This Kit is for the generation of template DNA using PCR without cloning process, providing all required reagents except the 1st primer set needed for the amplification of the target gene. The template DNA generated can be used to produce a target protein on manual procedure or on ExiProgen™.

Features and Benefits

Protein synthesis (expression and purification) without cloning within 1 day
Template DNA can be easily generated by just two step PCR from cDNA or genomic DNA in about 7 hours.
Therefore, target protein can be produced within a day using this kit with ExiProgen™.

Accurate protein template by high fidelity PCR
Because this kit uses AccuPower® ProFi Taq PCR premix and long & high fidelity PCR premix, you can obtain a
high quality template DNA with a minimal occurrence of error in the sequence.

High efficiency
This kit has been thoroughly optimized to ExiProgen™ so that small quantity of DNA can be used to produce up to 100 ug
of protein. (Optimal DNA quantity: A gene <1 Kb, 500 ng; A gene size of 1-2 Kb, 1 ug).


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Procedure

This kit uses two step PCR to generate template DNA. The procedure is described below.

1) 1st PCR: The DNA encoding a protein of interest will be amplified. The PCR product will contain start and stop codon, whole coding region of the protein, and additional sequences for the 2nd PCR. The primers for this step are gene specific primers with overhangs and can be ordered on the Bioneer's website.

2) 2nd PCR: Using a product from the 1st PCR, 2nd PCR performed using primer set by which T7 promoter and RBS are introduced at its 5’ end and T7 terminator is added at its 3' end. PCR product after 2nd PCR should be gel-purified for the cell-free protein synthesis.


 

Experimental results


Figure 1. Agarose gel data of template DNA synthesized by each PCR
Synthesis of linear template DNA with ExiProgen™ ProXpress PCR Template Kit. M1; 100bp DNA Ladder(Bioneer, D-1030), M2; 1kb DNA Ladder(Bioneer, D-1040) 1; SAV (Template – pT), 2; RNase H (Template – BL21(DE3) gDNA), 3; hGH (Template – pT) 4; CAT (Template - pBIVT), 5; UDG (Template - BL21(DE3) gDNA), 6; AcGFP (Template – pBIVT), 7; EVO (Template – pT), 8; RFP (Template – pIVEX), 9; Poly A polymerase (Template – pET15b),                


Figure 2. PAGE gel data for synthesized proteins.
Each linear template DNA generated by ExiProgen™ ProXpress PCR Template Kit are used as template for protein synthesis with ExiProgen™ EC Protein Synthesis Kit.
M; AccuLadder™ Protein Size Marker (Low)(Bioneer, D-2020), 1; SAV (13 kDa), 2; RNase H (20 kDa),3; hGH (23 kDa), 4; CAT (26.5 kDa), 5; UDG (28 kDa), 6; AcGFP (28 kDa),7; EVO (30 kDa), 8; RFP (31kDa), 9; Poly A polymerase (54 kDa)


 

Kit contents

Components K-7400 (16 reactions) K-7401(32 reactions)
AccuPower® ProFi Taq PCR Premix (20ul) 96 tubes 192 tubes
N terminus upstream cassette (5 ng/ul) 5 ul x 16 tubes (Yellow, NU) 5 ul x 32 tubes (Yellow, NU)
N terminus downstream cassette (5 ng/ul) 5 ul x 16 tubes (Yellow, ND) 5 ul x 32 tubes (Yellow, ND)
C terminus upstream cassette (5 ng/ul) 5 ul x 16 tubes (Violet, CU) 5 ul x 32 tubes (Violet, CU)
C terminus downstream cassette (5 ng/ul) 5 ul x 16 tubes (Violet, CD) 5 ul x 32 tubes (Violet, CD)
2nd Forward primer (10 pmoles/ul) 5 ul x 16 tubes (White, 2F) 5 ul x 32 tubes (White, 2F)
2nd Reverse primer (10 pmoles/ul) 5 ul x 16 tubes (White, 2R) 5 ul x 32 tubes (White, 2R)


 

Storage condition

at - 20°C

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Manual

Manual ExiProgen™ ProXpress PCR Template Kit


MSDS

MSDS ExiPrep™ ProXpress PCR Template Kit


Brochure

Brochure ExiProgen™ ProXpress PCR Template Kit


Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate

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