after bacterial transformation bacteria need to be speeded on a percitiplate to do so we need to pallet bacterial cell and it is subjected to centrifugation.
Generally cells that have just been transformed need treating very gently and would not be spun down until they have been plated and have formed colonies.16000 is much too high a G force unless spinning cells down to make dna so there is no need for the cells to survive. Why do you need to spin these cells at this stage?
Dear B R Siddharth
why do you centrifuge the cells after trasfomation? To reduce the volume and facilitate the cell spreading on the agar plates?
Generally i'm not perform centrifugation but i'm perform trasfromatino in 20ul of cells that are after shock resuspended in 200ul of LB or SOC and i plate the entire volulme. The rime required for the plating if depends also from the level of idratation of your plates. If your plates are wet, it require a long time, if you live it to get dry a little before plating, it will require less time.
IF you would like to plate, you have to do it sgort time and low speed and remove just a fraction of the liquid. (eg. 1500-2000g 5 minutes)
good luck
Manuele
@Paul Rutland actually i want to grow colonies of transformed cells with ligation products on my LB antibiotics plates thats why i spin down it so i had a doubt that how much spin i should give.
You can give single spin at 2700g after LB incubation of transformed cell and re-suspend in 100-180 microliter in same LB before plating.
In that case I would spin for 5 minutes at 1500g but would expect better survival if the cells were not spun at all
After incubating it at 37 degree Celsius for 1hr (shaking), centrifuge it at 8,000rpm for 3minutes. Discard the supernatant and then plate 200ul on the LB plate along with Antibiotics.
After incubating it at 37 degree Celsius for 1hr (shaking), centrifuge it at 8,000rpm for 3minutes. Discard the supernatant and then plate 200ul on the LB plate along with Antibiotics.
As Paul says, competent cells are delicate, and as others have pointed out, 11 000 g is not necessary. One thing you might try is to grow the cells in LB medium for a half hour before centrifuging, which would give them time to recover normal cell structure.
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