bacterial culture - (Nov/12/2013 )

hi guys,

i badly need your help. i had a sucessful cloning with insert in it a month ago. the clone was picked and grown in 5 ml lb with ampicillin overnight and then stored at 4 deg C. the originals plates are missing but i really need the same clone to grow again on a plate to be stored as i recently realised that is how to store the clone. so this lb with amp suspension that was sitting in 4 deg c had all the bacteria steled at bottom, i took 200 microltre and spread it on plate and left it in 37 deg c to grow overnight, am afraid all the bacteria are dead but i really need em. i was thinking, shoul i take some from this supension and try to grow in a suspension first and then plate the new suspension. or spin down this whole suspension and resuspend in new 100 microl lb and plate that or something. any suggfestions on how to get this growing will be greatly appreaciated.

cheers.

student47 on Tue Nov 12 10:04:31 2013 said:

hi guys,

i badly need your help. i had a sucessful cloning with insert in it a month ago. the clone was picked and grown in 5 ml lb with ampicillin overnight and then stored at 4 deg C. the originals plates are missing but i really need the same clone to grow again on a plate to be stored as i recently realised that is how to store the clone. so this lb with amp suspension that was sitting in 4 deg c had all the bacteria steled at bottom, i took 200 microltre and spread it on plate and left it in 37 deg c to grow overnight, am afraid all the bacteria are dead but i really need em. i was thinking, shoul i take some from this supension and try to grow in a suspension first and then plate the new suspension. or spin down this whole suspension and resuspend in new 100 microl lb and plate that or something. any suggfestions on how to get this growing will be greatly appreaciated.

cheers.

just wait and see what happened with the one you tested now...

If that failes, you can indeed do a liquid culture.

Just shake the flask a little bit (not too hard).

There should still be some live cells in there!

so i guess you mean take some from (how much?) the existing liquid cell culture and add it some lb amp and grow( around 5 ml lb with amp and grow at 37 deg with shaking o/n?)

you said dont shak too hard, so should i avoid spinning down too in case i was plaaning to use the whole amount of bacteria in there? may be i can use the whole existing culture into new 5 ml lb??

also does no shaking too hard means, i should grow them withou shaking??

very thanks for your advice already.

cheers