ExiProgen™



Bioneer’s ExiProgen™ is a breakthrough in synthetic biology allowing for the synthesis and purification of one to 16 proteins per run. DNA, in the form of plasmid or linear PCR product, is added to the system and Bioneer’s coupled transcription/translation E. coli extract then synthesizes the protein. The crude protein extract is further purified, using His-Tag engineered into the proteins. The net result is up to 100 ug of >90% pure Protein in about 6 hours. ExiProgen™ can also purify Nucleic Acids from several sources, making it one of the most versatile systems available.


Features and Benefits

  • Fast Protein production : From DNA to Pure protein in about 6 hours
  • DNA purification : A wide variety of gDNA kits available
  • RNA purification : A wide variety of RNA kits available
  • Affinity protein purification : For couples expression and purification in one system!

Built in Protocols optimized for protein synthesis/purification and the extraction of a wide variety of Nucleic Acid Samples
ExiProgen™ has more contains over 900 protocols, each optimized for protein synthesis/purification and target nucleic acid type and source sample. This optimization enables the user to obtain reproducible results for every run, every day. The instrument software can also be upgraded through the network connection port so you can stay up-to-date with the best performing protocols
Cooling Block
ExiProgen™ Has a built-in cooling block where the elution tube rack sits.
Sample integrity is ensured by keeping the samples below 10°C.
This allows for overnight runs and provides you with confidence in your results
Magnetic block & Heating block
To increase extraction efficiency, the ExiProgen™ has an integrated combined magnetic/heating block. The combination of bead magnetization and sample heating reduces experiment time and increases elution efficiency, resulting in increased sample yield. The heating block’s precision temperature control also ensures reproducible results for protein synthesis reactions
Contamination Shield
ExiProgen™ comes with a contamination shield designed to protect the assay from cross-contamination during instrument operation. Any time the pipette tips are moving, the contamination shield will slide under the tips, therefore eliminating the possibility of intra-assay cross-contamination which is a must when working with multiple samples
Easy to use with touch screen
The 3.5" touchscreen maximizes efficiency by offering an intuitive interface with simple push-button operation for processes such as selecting protocols and controlling the UV sterilization lamp.
UV lamp
ExiProgen™ has a powerful UV sterilization lamp that enables the user to sterilize the instrument chamber before and/or after every nucleic acid extraction or protein purification run. This prevents possible inter-assay cross-contamination that may occur on a busy work day


Experimental Procedure



Principle of protein synthesis and purification

ExiProgen™ Protein Synthesis Kit useds a E. coli extract to effect coupled transcription/translation of input DNA, which can be plasmid, or PCR generated DNA. The protein itself is generated with a His-Tag, which is then purified using the Ni-NTA magnetic bead provided. The result is high yields of protein that is >90% pure.



Nucleic acid extraction principle

ExiProgen™ DNA/ RNA Kits work on the principle of cell lysis, followed by bind, wash elute from silica magnetic beads. High yields of ultrapure DNA or RNA are obtained with OD260 readings of > 1.8 for DNA and 2.0 for RNA.





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Applications

Enzyme engineering, protein evolution, Synthetic biology, Bio-energy R&D, Protein drug R&D, expression screening, functional analysis, assay development and antigen production.


Specifications

Dimensions (W X H X D) 320 mm x 500 mm x 535 mm
Weight 27 kg
Operating temperature range 15 - 30°C
Operating humidity range 20 - 80%, no condensation
Operating system Built-in
User interface 320 x 240 touch screen graphic LCD
Input Voltage 100 - 240 VAC
Frequency 50 / 60 Hz
UV sterilization 5-minute cycle, 48.96 mJ/cm energy max 2
Communications TCP/IP
Heat block 40 - 95°C
Cooling block 4 - 10°C


Functions

  ExiProgen
DNA/RNA Extraction Yes
Protein Expression Yes
Protein Purification Yes
TFT Touch Screen Yes
Sample Number 16 samples
UV sterilization Yes
Heated magnetic block Yes
Contamination shield Yes
TCP/IP network connection Multiple systems can be daisy-chained together (up to 6)
Cooling system for sample storeage Yes
Automatic dispensing Yes
Power supply 24 VDC, 7.5 A(180 W)


Experimental Data

1. Protein expression and purification
- Amount of input DNA: 6 - 10 ug (purity >1.8)
- Protocol number : 902
- Total prep time : < 6 h
- Yield: ~100 ug


Procedure




SDS-PAGE analysis

Figure 1. GFP protein (His-tagged protein)’s expression/purification with ExiProgen™.
M; AccuLadder™ Protein Size Maker (Low)
E; expression sample
P; purification sample                   
Figure 2. Various His-tagged proteins expressed/purified with ExiProgen™.
M; AccuLadder™ Protein Size Maker (Low)
E; expression sample
P; purification sample                   


2. Total RNA Extraction from Tissue or cultured mammalian cell
- Sample: 15 ㎎ (Rat liver tissue) or 1 X 106 cells (HeLa)
- Protocol number: 202
- Average yield: 25 ug (Rat liver tissue), 10 ug (HeLa)
- Average RQS: > 9.5

* RQS(RNA quality score): he Caliper RQS is a calculated score that rates the quality of RNA samples. The RQS has been validated to correlate well with Agilent’s RIN (RNA Integrity Number) and follows the same 0-10 scale rating. Results comparing RIN to RQS for the same samples run on both LabChip GX and Agilent’s Bioanalyzer 2100 typically show <10% deviation. ( http://www.caliperls.jp/assets/pdf/lcgx/lcgx-AP-402.pdf)


Procedure



Rat total RNA Extracted from tissue

Total RNA from Rat was observed to contain intact rRNA with no evidence of degradation. 
 

Real-Time PCR carried out on Rat Total RNA
Real time RT-PCR was carried out.AccuPower® RocketScrpt™ RT PreMix(K-2101, Bioneer), and AccuPower® DualStar™ qPCR PreMix(K-6110, Bioneer) , and the Exicycler™ 96 Real-Time PCR system (A-2060).     


Analysis of total RNA from HeLa cells.

Total RNA was extracted from HeLa cell (1 X 106 cells) and analyzed using the LabChip GX (Caliper Life Science).
Each subunit of rRNA is clearly seen with no detectable degradation . In addition, the average yield of Total RNA was 10 ug, and the average RQS was 9.5 or higher.


Real-Time PCR carried out on total RNA from Hela cells.

Real time RT-PCR was carried out on total RNA extracted from HeLa cell (1 X 106 cells) using AccuPower® RocketScrpt™ RT PreMix(K-2101, Bioneer), AccuPower® DualStar™ qPCR PreMix(K-6110, Bioneer) and the Exicycler™ 96 Real-Time PCR system(A-2060)

 

3. Genomic DNA from Cultured mammalian cell (HeLa))
- Sample volume: 1 X 106 cells
- Protocol number: 109
- Total prep time: < 1hr
- Average yield: 4-8 ug


Procedure



Agarose gel analysisAgarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 1X106 cells of E.coli cells and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH2O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

4. Genomic DNA from Tissue (Bovine skeletal muscle)
- Sample volume: 1 X 106 cells
- Protocol number: 109
- Total prep time: < 1hr
- Average yield: 8-12 ug


Procedure


Agarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 20 mg of bovine tissues and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH2O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

5. Genomic DNA from Plant Tissue (Spinach leaf)
- Sample volume: 100 ㎎
- Protocol number: 104
- Total prep time: <1 hr
- Average yield: 3-5 ug


Procedure



Agarose gel analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 100 mg of spinach leaves and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH2O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 100 mg of spinach leaves and Lanes 2, 4, 6, 8, 10, 12, 14, 16 were extracted with ddH2O as a negative control in DNA extraction. Note all samples have very similar yields.
Purity was also tested and was consistently between 1.9 and 2.0 (not shown).

 

 




 

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Application Note

Protein Synthesis

ExiProgen™_Application_Fluorescent proteins

ExiProgen™_Application_High molecular weight protein

ExiProgen™_Application_in vivo protein expression pET vectors


Manual

ExiProgen™


Brochure

ExiProgen™


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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