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Resuspending bacterial pellet to a certain OD600

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Resuspending bacterial pellet to a certain OD600

Postby biology_06er » Fri Jun 29, 2012 2:23 am

Hi there,

I plan to do some flow next week and an one of the steps is to resuspend the bacterial pellet to an OD600 0.5. So say I had a 1.5mL overnight culture and the following morning the OD600 was 1.7. Once I spin down, wash etc etc I read to get to a certain OD (0.5 in my case)...this is the calculation


Divide the OD you want (0.5) by the OD bacteria was at in morning (1.7)=0.5/1.7=0.29=giving you a concentration factor (cf)

Divide the original culture volume (1.5mL) by the cf

1.5/0.29=5.1mL


Ok..so I don't need 5.1mL of culture to subsequently use..say altogether I only need 2mL how do I go about this? So that in the end I have 2mL of bacteria with an OD of 0.5?


Man, I hope I make sense!

Cheers,

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Postby JackBean » Fri Jun 29, 2012 7:12 am

What about to get only aliquot of your O/N culture? :roll:
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Re: Resuspending bacterial pellet to a certain OD600

Postby biology_06er » Fri Jun 29, 2012 10:00 am

As in from my o.n culture take 0.5/1.7=0.29*2mL=0.588..So do I take 588uL of my overnight culture..spin it down wash etc etc..then resuspend the pellet in 2mL?
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Postby JackBean » Fri Jun 29, 2012 10:17 am

something like that
http://www.biolib.cz/en/main/

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Re: Resuspending bacterial pellet to a certain OD600

Postby biology_06er » Fri Jun 29, 2012 9:54 pm

Something like that?..Which means a part is wrong?
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Postby biology_06er » Fri Jun 29, 2012 10:08 pm

That sounds wrong..

Is this way better..say I had the a 10mL of o/n starter culture
I take 1mL from that starter culture, wash, spin, block etc..and when it comes down to resuspending my pellet..i resuspend in 1.5mL again..therefore I technically have a culture at an OD600 of 1.7.

From that culture which I just resuspended in...can I take 441uL and then top up 2mL using 1559uL of my buffer?
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Postby JackBean » Sun Jul 01, 2012 9:53 am

If you take 1 ml and dilute it into 1.5 ml, you will hardly have the same OD :roll:
http://www.biolib.cz/en/main/

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Re: Resuspending bacterial pellet to a certain OD600

Postby biology_06er » Sun Jul 01, 2012 11:53 am

Urg..I meant resuspened in 1mL
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Postby biology_06er » Sun Jul 01, 2012 11:55 am

and from that 1mL resuspended culture..I take

0.5*2mL/1.7=588uL and top up to 2mL
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