After isolation of plasmid i diluted it by adding 500 ul of sterile 1X TE to get the stock of 1000 ng/ul. I want to concentrate the solution. Is there any method available for this ?
Dear Prakash,
You may use the procedure contained in the following link:
http://arbl.cvmbs.colostate.edu/hbooks/genetics/biotech/manip/conc.html
Hoping this will be helpful,
Rafik
Dear Prakash, the simple way to concentrate the plasmid solution is to freeze dry the sample and resuspend in required volume.
Hi Prakash,
It depends on how much you want to concentrate the plasmid. You have to keep in mind that freeze drying will also concentrate the TE buffer. A 10x TE can be a problem for downstream experiments. So if you want to concentrate the plasmid 10X it is better to precipitate. If you just need a 2x concentration you can use freeze drying or simply put it in a speed vac.
Dear
salts can be concentrated diluted Plasmid. You could reduce the volume by extraction with butanol.
Thank you
Marius
Dear Rafik,
Thank you for your valuable suggestion. After following the above link i got the result. But it needs little modification i.e. the 3 M sodium acetate must be pre-chilled prior to use.
Dear
Add equal volume of 100% absolute ethanol, spin the sample in vacuum centrifuge for half hour, then allow the ethanol to evaporate by opening the lid of the eppendorf tube, after that add the desired volume of water or EB buffer to get a concentrated sample of your plasmid.
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