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#16 ascacioc

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Posted 12 September 2012 - 05:29 AM

@phage: this is why I love this forum: you always find new ways others do stuff: nice beaker idea, better than my sticky tape that does not always stick, especially when you have 20 tubes :D

@Sameer: while waiting for me to check all the cloning strategy from the start you can retry transformation as phage says: with SOC and good shaking, this only if you have ligation reaction left or stuff with which you can repeat the ligation reaction quickly tomorrow. Today prepare SOC media and autoclave it :)

Andreea

#17 siddharthsameer

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Posted 12 September 2012 - 05:43 AM

@phage: this is why I love this forum: you always find new ways others do stuff: nice beaker idea, better than my sticky tape that does not always stick, especially when you have 20 tubes Posted Image

@Sameer: while waiting for me to check all the cloning strategy from the start you can retry transformation as phage says: with SOC and good shaking, this only if you have ligation reaction left or stuff with which you can repeat the ligation reaction quickly tomorrow. Today prepare SOC media and autoclave it Posted Image

Andreea

@ andrea my supervisor asked me to wait until tommorow to see the colony, and i am mere a master student so cant say anything to her right now :(...rest of the episode about my lovey supervisor tell u später :)

#18 ascacioc

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Posted 12 September 2012 - 05:53 AM

what about doing like she said (which is so not gonna work; i tell you from now that nothing besides a contamination would grow on that plate until tomorrow) and prepare SOC media for next time :) I mean anyhow you need to autoclave it and aliquot it (in autoclaved tubes of course) and since basically your task is waiting right now, at least wait while doing something useful :P

#19 phage434

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Posted 12 September 2012 - 07:02 AM

You might also want to re-make your competent cells, or at least grow up a 10 ml overnight culture to be prepared to inoculate a 1 liter culture tomorrow.

For the SOC, you could aliquot first and then autoclave the aliquots in bulk. One less chance to contaminate things.

#20 ascacioc

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Posted 12 September 2012 - 07:08 AM

@phage: don't you first do SOB and then add glucose and Mg salts after autoclaving them separately? I thought glucose in media and autoclave are not a happy mixture.

#21 phage434

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Posted 12 September 2012 - 07:03 PM

Good point. Yes, your method is better.




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