Bio-rad MicroPulser™ Electroporator User Manual

MicroPulser™Electroporation ApparatusOperating Instructionsand Applications GuideCatalog Number165-2100For Technical Service Call Your Local Bio-Rad O

at 5 milliseconds when working with high-resistance samples. For these organisms, theMicroPulser has pre-programmed settings for delivery of the corre

transformants recovered is the product of the transformation frequency and the number ofcells present, the transformation efficiency (transformants/µg

Fig. 4. Resistance of solutions of (A) NaCl and MgCl2 and of (B) buffers of NaPO4 at pH 6.1 and 7.3and HEPES at pH 7.5. Resistance was measured in 0.2

Fig. 5. MicroPulser control panel.Section 4MicroPulser Operating Instructions Refer to Figure 1 for a view of the components of the MicroPulser system

StAS. aureus (0,2 cm cuvette) 1.8 1 2.5AgrA. tumefaciens(0.1 cm cuvette) 2.2 1 -Ec3E. coli(0.2 cm cuvette) 3.0 1 -Included under the Fungi Settings pr

3. Pulse FunctionPressing the "Pulse" button results in the capacitor charging to the set voltage; during thistime "PLS" is shown

Section 5High Efficiency Electrotransformation of E. coliElectroporation provides a method of transforming E. coli at efficiencies as high as 109to 10

4. Transfer the mixture of cells and DNA to a cold electroporation cuvette and tap thesuspension to the bottom. Place the cuvette in the chamber slide

7. Resuspend the cell pellet in 25 ml of sterile, ice-cold 10% glycerol. Transfer to a 30 mlsterile Oakridge tube. Pellet the cells by centrifugation

Section 7Electroporation of Agrobacterium tumefaciens7 .1 Preparation of Electrocompetent CellsSee Lin (1995) for additional information.1. Inoculate

Table of ContentsPageSection 1 Safety Information ...11.1 Electri

8. Incubate the cells 3 hr at 30 °C, shaking at 250 rpm. Plate aliquots of the electroporatedcells on YM agar plates containing the appropriate select

4. Transfer the DNA-cell samples to the appropriate electroporation cuvettes that have beenchilled in ice and tap the suspension to the bottom of the

9.2 Electroporation1. Pipette the DNA samples (up to 1 µg) to be electroporated into sterile 1.5 ml microfugetubes. Place tubes on ice.2. Add 200 µl

10.2 Electroporation1. Pipette the DNA samples (up to 50 µg) to be electroporated into sterile 1.5 ml microfugetubes. Place tubes on ice.2. Add 800 µl

5. Add 50 ml of sterile YPD/HEPES to each of the bottles and vortex to resuspend the cellpellets; add 1.25 ml of 1M DTT to each bottle; mix gently. In

Appendix IReferences1. Allen, S.P. and Blaschek, H.P., Factors involved in the electroporation-induced transformation ofClostridium perfringens, Appl.

25. Siguret, V., Ribba, A.-S., Cherel, G., Meyer, D., and Pieru, G., Effect of plasmid size on transfor-mation efficiency by electrtoporation of Esche

Electrical Problem Possible cause and solutionVerify the display LED shows a reading between “5.50” and “6.50”.If problems persists, contact Bio-Rad.1

BiologicalThe general symptom addressed in this section is transformation efficiencies that are toolow to detect or too low to be useful. The followin

Appendix IIIProduct InformationSpecificationsInput voltage 100–120 V RMS, 50/60 Hz;automatic mains voltageswitching220–240 V RMS, 50/60 HzInput curren

Appendix I References ...21Appendix II Troubleshooting

CatalogNumber Product DescriptionPlasmid purification732-6100 Quantum Prep Miniprep Kit, 100 preps, includes 20 ml cellresuspension buffer, 25 ml cell

Life ScienceGroupWebsite www.bio-rad.com Bio-Rad Laboratories Main Office 2000 Alfred Nobel Drive, Hercules, CA 94547, Ph. (510) 741-1000, Fx. (510)7

WarrantyBio-Rad Laboratories warrants the MicroPulser against defects in materials and workmanship for1 year. If any defects occur in the instrument d

Section 1Safety InformationRead This Information Carefully Before Using The MicroPulser.The MicroPulser meets the safety requirements of EN61010 and t

2Warning: The MicroPulser generates, uses, and radiates radio frequency energy. If it is notused in accordance with the instructions given in this man

The capacitance discharge circuit of the MicroPulser generates an electrical pulse with anexponential decay waveform (Figure 2). When the capacitor is

The purpose of the 30 ohm series resistor in the MicroPulser is to protect the instrumentcircuitry should arcing occur. Under normal operation, when s

The MicroPulser also includes a means to truncate the exponential decay pulse soonerthan the expected time constant as long as the voltage is greater