商品介紹

ELEPO21   In Vitro High Energy Electroporator 泛用型電穿孔儀

        Transformation (轉形) of

                  - Bacterial cells

                  - Fungi, Yeasts, etc.

 

        Transfection (轉染) of

                  - Primary Cell Cultures

                   - Any Cell Lines ( animal cells and plant cells)

                   - Stem Cells, ES Cells, iPS Cells, etc.

 

 

ELEPO21 神奇的四階段多脈衝設計

神奇的四階段脈衝搭配電壓衰減, 達到高 transformation / transfection 效率及高存活率.

1) Poring Pulse Mode:

    較高的電壓,較短脈衝時間, 多脈衝及電壓衰減. 此階段是在產生最低傷害下於細胞膜形成小孔洞.

2) Polarity Exchanged Poring Pulse

   打孔脈衝的極性反轉.

3) Transfer Pulse Mode:

    較低的電壓,較長脈衝時間, 多脈衝及電壓衰減. 此階段是在產生最低傷害下將 target molecules (DNA, RNA, etc.)導入細胞.

4) Polarity Exchanged Transfer Pulse

    導入脈衝的極性反轉, 提高 transformation / transfection 效率.

 

Hi Voltage Electroporation

ELEPO21 In Vitro 高電壓電穿孔儀是由Nepa Gene公司開發的獨特脈衝系統, 是由4階段多脈衝搭配電壓衰減的設計,與傳統的單一指數波衰減的電穿孔儀比較, 對 bacteria, yeasts, and fungi 的transformation效率有非常顯著的提高.

 

High-effciency gene transfer in bacteria by multi-step electroporation

- Transformation data of bacteria (E. coli) -

We measured gene transfer efficiency using the ELEPO21 in Gram-negative bacteria. Competent cells were prepared as usual from E. coli DH5α. The competent cells were mixed with pUC19 DNA, and a 20μl aliquot (containing 10^9-10^11 cells and 10 pg DNA in 10％ glycerol solution) was transferred to the 1 mm gap electrode cuvette (EC-001S, Nepa Gene). The cuvette was set in the chamber connected to the ELEPO21, and delivered 3-step pulses as described below. All steps were done on ice. After electroporation, the cells were plated on LB agarose medium containing ampicillin, and colonies formed were counted. Transformation efficiency was expressed as a number of colonies per ug plasmid DNA used.

[ELEPO21　pulsing conditions, Fig. 1]

Poring Pulse (voltage: 2,000 V, pulse length: 2.5 msec, pulse interval: 50 ms, number of pulses: 1, polarity: +)

Transfer Pulse (voltage: 150 V, pulse length: 50 msec, pulse interval: 50 ms, number of pulses: 5, polarity: +/-) 

To evaluate the above results, we measured gene transfer efficiencies using a conventional electroporator (ECM630, BTX) that deliver a single exponential pulse as described below. 

 

[ECM630 pulsing conditions, Fig. 2]

Single pulse (voltage: 2,000 V, resistance: 200 ohms, capacitance: 25 uF, number of pulse: 1)

 

Fig. 1: ELEPO21 pulse shape	Fig. 2: ECM630 pulse shape

Fig. 3

實驗結果:

The above cell suspensions (sample resistance value: 7.7 K ohms) were used for electroporation. The tranformation efficiency obtained by the ELEPO21 electroporator was approximately 5 times higher than that by the ECM630 electroporator (Fig. 3).

*The values are averages of repeated experiments.

*The optimum pulsing conditions were used for ELEPO21 and ECM630.

*cfu: colony forming unit. 

 

High-effciency gene transfer in Yeast by multi-step electroporation

- Transformation data of Yeast (S. cerevisiae) -

We measured gene transfer efficiency using the ELEPO21 in yeast. Competent cells were prepared as usual from budding yeast S. cerevisiae. The competent cells were mixed with pAS2 DNA, and a 20μl aliquot (containing 108-1010 cells and 50 ng DNA in 1M sorbitol solution) was transferred to the 1 mm gap electrode cuvette (EC-001, Nepa Gene). The cuvette was set in the chamber connected to the ELEPO21, and delivered 3-step pulses as described below. All steps were done on ice. After electroporation, the cells were plated on selective agarose medium devoid of nutrients, and the colonies formed were counted. Transformation efficiency was expessed as a number of colonieis per μg plasmid DNA.

[ELEPO21　pulsing conditions, Fig. 1]

Poring Pulse（voltage: 500 V, pulse length：1.5 msec, pulse interval：50 ms, number of pulses：5, polarity：+）

Transfer Pulse（voltage: 50 V, pulse length：50 msec, pulse interval：50 ms, number of pulses：5, polarity：+/-） 

To evaluate the above results, we measured gene transfer efficiencies using a conventional electroporator (ECM630, BTX) that deliver a single exponential pulse as described below. 

 

[ECM630 pulsing conditions, Fig. 2]

Single pulse (voltage：750 V, resistance：200Ω, capacitance：25μF, number of pulse：1)

Fig. 1: ELEPO21	Fig. 2: ECM630

Fig. 3

實驗結果:

The above cell suspensions (sample resistance value：12.36 KΩ) were used for electroporation. The tranformation efficiency obtained by the ELEPO21 was approximately 6 times higher than that by the ECM630 electroporator (Fig. 1-3).

*The values are averages of repeated experiments.

*The optimum pulsing conditions were used for ELEPO21 and ECM630.

*cfu: colony forming unit. 

 

Low Voltage Electroporation

ELEPO21 對很難轉染的細胞例如 primary cells, stem cells, immune cells, blood cells 等 的transfection效率與存活率都很高, 而且不需要特殊的耗材與buffer.

 

Transfection into Primary Cells

 

BMMC Primary Mouse Bone Marrow-Derived cells

存活率: 61%	轉染效率: 68%

FACS Data

 

 

 

Transfection into Stem cells, ES cells 及 iPS cells