I made a stupid mistake and need urgent help

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I made a stupid mistake and need urgent help

Post by moni21 » Tue Apr 10, 2012 1:06 am

Hi Everyone,

I made a terrible, terrible mistake and I need your help. While I was doing minipreps (isolating my plasmid DNA with the insert in it) I threw away all of the bacterial pellets. However, I did save 0.5 ml of the culture (LB+amp and the specific colony). How should I re-grow my colonies? I mean do I need to re-plate them or can I just use a small amount of the liquid that I saved to grow overnight in LB broth? Also, does freezing your bacteria without glycerol kill it? I know I should know all these things and I'm really sorry, but I'm new to this. Worst case scenario, I would have to repick all of my colonies and do everything all over again, which I could do, but I just don't have the time.
Help please!!!


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Post by JackBean » Tue Apr 10, 2012 7:39 am

Just use your liquid culture to grow it again. If you couldn't do it, you've had to pick new colonies every time :lol: ;)

Did you at least throw them into nitrogen or just put them into fridge? You can try to grow them, but I would use higher amount than usual.

Cis or trans? That's what matters.

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