By Jac A. Nickoloff
Harvard college, Boston, Massachusetts. handbook of protocols within the electroporation of proteins and DNA into fish, insect, and mammalian cells. For researchers. Plastic comb spiral binding. sixty five members, 28 U.S.
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Additional resources for Animal Cell Electroporation and Electrofusion Protocols (Methods in Molecular Biology)
Using a pipet, drop the metaphase cells onto clean glass microscope slides and allow them to air-dry. 8. Stain the slides with 4% Giemsa for 2-5 min, dry, and permanently mount them with a covershp for microscopic analysis of cytogenetic abnormalities. Morgan 68 and Day Table 1 Chromosomal Aberration Yields, Platmg Efficiency, and Cell Surwval m CHO Cells After Exposure to PvuII (Adapted from Yates et al. 5 a Number of cells showing 2 1 chromosomal rearrangement. Each pomt represents the mean of two independent experrments One hundred metaphase spreads were analyzed for each experrment.
Table 6 shows a survey of commercially available chambers. 6. Measuring ECM Parameters By following an established protocol, it is generally not necessary to measure the ECM parameters, especially if the same types of generator Instrumentation 57 Fig. 9. Petri dish electrodes for large-volume electroporation of mammalian cells in suspension or adherence, and electroporation of fish eggs. and chamber are used (different generators might give different output voltages for the same charging voltage setting).
Meander-type chamber for visual observation of fusion. with PBS. The parallel plate electrode configuration in a Petri dish is also very useful for electroporation of adherent cells, if the electrodes are situated so they touch the Petri dish bottom. Instead of parallel plates, an array of concentric electrodes can also be used to create a large-volume ECM chamber in a Petri dish (II). 3. Small-Volume Flowthrough Chambers for the ECM of Large Volumes If it is required to transform large volumes (above 50 mL), it is economical to pulse the generator repetitively in synchrony with a pump that pushes the medium with the cells and transformants through a relatively small chamber.
Animal Cell Electroporation and Electrofusion Protocols (Methods in Molecular Biology) by Jac A. Nickoloff