Electroporation Cuvettes, 0.1 cm - FAQs

当使用高电压在导电缓冲液中进行电转时，可能会出现电火花。MgCl2和PO4的导电能力尤其强。

以下为一些建议：
1.将克隆反应的离子强度保持在最小。如果反应缓冲液含有高盐，在电转前请将DNA样本用水或TE缓冲液稀释。
2.尽量避免加入导电离子。DNA溶液的体积不应超过反应总体积的5%。例如：40 μl细胞中加入2 μl DNA。
3.确保没有气泡存在。
4.确保电极接触是干净紧密的。擦除电转杯外面的任何沉淀物。
5.要获得最佳结果，细胞应当被加入缝隙的最底部。轻弹电转杯使细胞沉降到底部。

Our cuvettes are known as the "Potter-style" cuvettes, and they fit most electroporators that accept a standard size cuvette.

When electroporating cells at high voltage in conductive buffers, arcing may occur. MgCl₂ and PO₄ in particular are very conductive.

Some suggestions:

1) Keep ionic strength of cloning reactions to a minimum. If reaction buffers contain high salt, dilute DNA sample in water or TE buffer before electroporation.

2) Minimize addition of conductive ions. The volume of DNA solution should not exceed 5% of the total reaction. Example: 2 ?l DNA per 40 ?l of cells.

3) Be sure no air bubbles are present.

4) Make sure the electrical contacts are clean and tight. Wipe away any condensation on the outside of the cuvette.

5) For best results, the cells should be aliquoted into the bottom of the gap - tap the cuvette gently to help the cells settle to the bottom.

The two most important electrical parameters for consideration in electroporation are pulse length and field strength. Field strength is defined as volts/centimeter, where V is equal to the initial peak voltage and cm is equal to the measurement of the gap between the electrodes of the cuvette in centimeters. For example, during electroporation of bacteria at 1,500 Volts in a 0.1 cm cuvette, the field strength would be 15,000 volts/cm, or 15 kV/cm. Typically, electroporation of bacteria requires field strengths of greater than 15 kV/cm. Yeast cells require 6-8 kV/cm, and mammalian cells often require optimization between 0.5 to 2.5 kV/cm.

20-80 ul is a general suggested volume, but follow the optimized protocols for the cells and cuvettes that you are using.