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Overview

PUREfrex® is an ultra-cool, ultra-smart, and ultra-useful in vitro coupled transcription/translation system based on E.coli translation machinery but completely different from S30 translation extracts. PUREfrex® is NOT AN EXTRACT.  PUREfrex® is a RECONSTITUTED SYSTEM comprising highly purified E. coli ribosomes, highly purified recombinant proteins, and other highly purified factors required for transcription, translation, aminoacylation, and energy regeneration. As such, PUREfrex kits provide previously unthinkable reaction controllability compared to cell extract-based systems and a well defined mix of molecular components from which to purify synthesized protein. 

Purefrex 2.0 Ordering Information
Product Name	Product Number	Total Reaction Volume per Kit
PUREfrex 2.0	GFK-PF201-0.25-EX	250 ul
	GFK-PF201-0.25-5-EX	5 x 250 ul (packaged as 5 x 250 ul) Sales to be discontinued
Standard kit for use when protein yield is the primary goal and disulfide bonding is not required

Which PUREfrex Kit Is Right For Me?

PUREfrex® coupled transcription/translation system kits are available in four versions. 

• PUREfrex® 2.0 - Choose 2.0 when protein yield is you primary goal and disulfide bonding is not relevant.
• PUREfrex® 2.1 -  2.1 kits are based on 2.0 reaction components but are not premixed with reducing agent. 2.1 kits include separate vials of cysteine, DTT, and reduced glutathione allowing control of the redox environment. Choose 2.1 kits for primarily for target proteins with disulfide bonds.   
• PUREfrex® 1.0 - 1.0 kits produce less protein than 2.X kits, but the complete contents of 1.0 kits is disclosed. Chose 1.0 when complete information on PUREfrex reaction components takes precedence over protein yield. 
• PUREfrex® 1.0 Custom - Gene Fronteir is happy to provide custom reaction mixtures whenever feasible. Custom PUREfrex kits are based on 1.0 reaction components. Please contact us with your request.  

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For detailed information about each PUREfrex® version, please use the navigation links in the sidebar at left. 

Please Note: All protein components in all PUREfrex versions are tag free allowing the synthesis of protein with any desired tag.  

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About PUREfrex 2.0

PUREfrex® 1.0 was introduced in 2011. Extensive R&D efforts by Gene Frontier led to the release of PUREfrex® 2.0 in 2015, with 2-10 times the protein productivity of 1.0, fewer impurities, and other desirable performance characteristics.  

2.0 vs 1.0 Performance

 Protein yields with version 2.0 are generally significantly higher than with version 1.0 but the reaction components of v1.0 are more fully disclosed (see "components" table below). Version 1.0 is therefore more appropriate when the most precisely defined system is desirable.

Purefrex Version Performance Comparison
PUREfrex Version	1.0	2.0/2.1
Specification
Yield of synthesized DHFR	150 ug/ml	600 ug/ml
Reaction volume required for 100ug DHFR	660 ul (> 3 kits)	160 ul (< 1 kit)
Yield of synthesized GFP	100 ug/ml	800 ug/ml
Reaction volume required for 100ug GFP	1000 ul (> 4 kits)	125 ul (< 1 kit)
Version Selection Guidance
Protein Preparation	--	++
Protein Science (translation, protein folding, etc.)	+	--
Synthetic Biology	+	+

 The SDS-PAGE gel below compares performance between 1.0 and 2.0 for several proteins. 1 ul of a standard 20ul reaction mixture was applied to each lane following a 4 hour reaction. 

 

PUREfrex v2.0 has less contaminants 

Improved purification of PUREfrex® v2.0 components greatly reduced the levels of RNase and ß-galactosidase contaminants. In addition, lipopolysaccharide (LPS) levels are reduced to to approximately 0.1 EU per 1 µL of reaction mixture. All proteinous components of PUREfrex®2.0 have no tags for purification and detection. It allows to fuse your protein with any tag to purify the product.

 

PUREfrex 2.0 Kit Contents and Storage
Reagent	Quantity	Description	Storage
Solution I	125 µL	Amino acids, NTPs, tRNAs and substrates for enzymes etc.	-20 °C
Solution II	12.5 µL	Proteins in 30 % glycerol buffer	-20 °C or -80 °C^2
Solution III	25 µL	Ribosome (20 µM)	-80 °C^2
DHFR DNA ^3	10 µL	PCR product containing a gene encoding dihydrofolate reductase (DHFR) from E.coli as a positive control. (20 ng/µL)	-20 °C
Indicated temperature shows storage temperature after opening the kit. Please store the kit at -80 °C before opening. The rest of solution should be frozen rapidly in liquid nitrogen or dry ice/ethanol, and be stored at -80 °C. Divide into aliquots, if necessary, and avoid refreeze and thaw as much as possible. For DHFR synthesis, use 1 µL of DHFR DNA for 20 µL of reaction. DHFR DNA sequence.

PUREfrex 2.0 General Reaction Protocol

1) The amounts given are for 20 µL reaction. For scaling up the reaction, adjust the volume of reagents accordingly.
2) Thaw Solution I by incubation at room temperature or 37 °C for 1 minute for completely dissolving, and then cool on ice.
3) Thaw Solution II and III on ice.
4) Mix Solution I, II and III respectively by vortex and centrifuge briefly to collect each solution at the bottom.
5) Assemble the reaction mixture in a tube as follows. Add the template DNA to 1 – 3 ng/µL per 1 kb.
6) Incubate the tube at 37 °C for 2 – 4 hours on a heat block or a water bath.
7) Analyze the synthesized products. Please add the same amount of H2O to the reaction for the sample of SDS-PAGE.

Contents of a standard PUREfrex reaction	volume
Water	7-X µL
Solution I	10 µL
Solution II	1 µL
Solution III	1 µL
Template DNA *	X µL
Total	20 µL
* Please visit the template DNA preparation site

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PUREfrex Ordering Information
Product Name (click for price and order info)	Catalog Number	Total Reaction Volume	Notes	Product Information Page
Kits
PUREfrex 2.0	GFK-PF201-0.25-EX	250 ul		2.0
	GFK-PF201-0.25-5-EX	5 x 250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF201-2ML-EX	1 x 2 ml	New size !
	GFK-PF201-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF201-10-EX
Current generation kit for protein synthesis. Contains DTT as a reducing agent.
PUREfrex 2.1	GFK-PF213-0.25-EX	250 ul		2.1
	GFK-PF213-0.25-5-EX	5 x 250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF213-2ML-EX	1 x 2 ml	New size!
	GFK-PF213-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF213-10-EX
Current generation kit for proteins with disulfide bonds. Formulated for user control of redox environment.
PUREfrex 1.0	GFK-PF001-0.25-EX	250 ul		1.0
	GFK-PF001-0.25-5-EX	1250 ul (packaged as 5 x 250 ul)	Sales to be discontinued
	GFK-PF001-2ML-EX	1 x 2 ml	New size!
	GFK-PF001-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF001-10-EX
Prior generation kit for protein science research and synthetic biology. Components are fully disclosed. Custom formulated kits are based on 1.0 components. Please inquire about custom kits.
PUREfrex Reaction Supplements
DnaK Mix	GFK-PF003-0.5-EX	for 500 ul of reaction mix		Supplements
	GFK-PF003-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF003-10-EX
GroE Mix	GFK-PF004-0.5-EX	for 500 ul of reaction mix		Supplements
	PF004-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF004-10-EX
Supplements for synthesis of aggregate-prone proteins.
DsbC Set	GFK-PF005-0.5-EX	for 500 ul of reaction mix		Supplements
	PF005-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF005-10-EX
PDI Set	GFK-PF006-0.5-EX	for 500 ul of reaction mix		Supplements
	PF006-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PF006-10-EX
Supplements for synthesis of proteins containing disufide bonds.
EF-P	GFK-PFS052-0.5-EX	for 500 ul of reaction mix		Supplements
	PFS052-10ML-EX	5 x 2 ml	Note: cat. no. change from GFK-PFS052-10-EX
Supplement for use when consecutive prolines are in target protein.

PUREfrex® is for in vitro research only. PUREfrex® should not be used for the therapy, diagnostic or administration to animals including human and should not be used as food or cosmetics etc. To avoid the contamination of nuclease, nuclease-free-treated water, reagents and materials should be used. We also recommend wearing gloves and mask.

"PUREfrex" is a registered trademark of the Gene Frontier Corporation. All rights reserved. 2021.