ExiProgen™ :
Synthetic Biology meets Automation Technology

The field of biology and many commercial industries are directing their attention to synthetic biology to solve increasingly complex questions and problems. Commonly used methods to achieve synthetic biology goals are cumbersome and slow at best. But that’s changed thanks to Bioneer’s ExiProgen™ system! ExiProgen™ is a breakthrough in synthetic biology allowing for the synthesis and purification of one to 16 proteins per run. Simply add DNA, press run and in about 6 hours the system will synthesize and purify up to 16 different proteins. 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

Fully automatic: DNA-in-Protein-out:	Pipette 6~10 μg of DNA into the kit and load into ExiProgen for high-purity proteins within 6 hours.
Parallel Process:	Synthesize and purify1 to 16 unique proteins simultaneously.
High performance:	The system enables synthesis of proteins that are toxic to in vivo expression systems.
High purity and High yield:	Each well will yield up to 100 μg protein that is over 90% pure.
Reproducibility:	Since you only need to add template DNA to obtain your protein, reproducibility is maximized.

Function

Feature	Benefit
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	Sample preparation for bio-defense detection of various bacteria and viruses

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
Temperature range	15 - 30°C
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

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 analysis

Lanes 1, 3, 5, 7, 9, 11, 13, 15 were extracted with 1X10⁶ cells of cultured HeLa cell 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 1.8 and 2.0 (not shown).

4. Genomic DNA from Bacteria (E.coli)
- Sample volume: 1 X 10⁶ cells
- Protocol number: 109
- Total prep time: < 1hr
- Average yield: 4-8 ug

Procedure

Agarose 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).

5. 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).

6. 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).