A high-salinity solution with calcium chloride enables RNase-free, easy plasmid isolation within 55 minutes

Sasagawa, Noboru; Koebis, Michinori; Yonemura, Yoji; Mitsuhashi, Hiroaki; Ishiura, Shoichi

doi:10.5582/bst.2013.v7.6.270

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“…Based on the alkaline lysis method, we developed a new plasmid purification method, which ends within 1 hour and does not need RNase (Figure 5) [22]. The principle of this method is a combination of the alkaline lysis method, CaCl 2 precipitation, and PEG precipitation.…”

Section: -Minute Methodsmentioning

confidence: 99%

Plasmid Purification

Sasagawa¹

2019

Plasmid

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Plasmid purification is a rather classical experiment, but the technique is still developing for time-and cost-saving. The critical principle is based on the alkaline lysis method, although the following steps have several variations. The needed purities and/or quantities of DNA depend on researches using isolated plasmids, meaning that more reasonable method can be selected in each experiment. For example, a non-alkaline-lysis method such as boiling method is still available. One of the important steps for purifying plasmid is a removal of RNA. Ribonuclease is usually used for removing RNA from plasmid sample. On the other hand, a kind of salts such as lithium and calcium functions to make RNA as a selective precipitate from DNA-RNA mixture. Based on these backgrounds, the technique to purify plasmid DNA has been discussed.

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Section: -Minute Methodsmentioning

confidence: 99%