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OD range for spreading on agar

microbiology general techniques plating

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5 replies to this topic

#1 Omnimax

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Posted 23 August 2012 - 07:24 AM

Hi all,

I want to resuspend colonies from a mixed culture (environmental, no strains) that I have on a plate (M9 agar) and respread it on a new plate to do a little comparison on how well they will grow on different solid media. Problem is I am not sure how much (both in volume and OD) would normally be ok for spreading on a plate in order to avoid a mess of growth or an empty plate. Is there a range of "suitable" ODs for plating?

Thanks in advance,
Omni

#2 phage434

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Posted 23 August 2012 - 08:13 AM

OD 1 is very approximately 10^9 cells/ml (varying a lot by species and medium, and phase of moon). You probably don't want more than a thousand cells on your plate, so you want to dilute 10^5 (adjust for measured OD) and then plate 100 ul. You'll likely hit the right place if you did this with 10^4, 10^5, and 10^6 dilutions of an OD 1 culture.

#3 Omnimax

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Posted 23 August 2012 - 01:24 PM

Thanks a lot! I'll try this tomorrow and see how it works out.

#4 pito

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Posted 24 August 2012 - 06:26 AM

THe use of OD is something that is being misused by many.
I would advise you to check the attachements.
Try to understand it , if you do, it will make everything a lot easier and you would not make mistakes like so many others (even in published papers) do.

Attached Files


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#5 lyok

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Posted 26 August 2012 - 09:41 AM

OD 1 is very approximately 10^9 cells/ml (varying a lot by species and medium, and phase of moon). You probably don't want more than a thousand cells on your plate, so you want to dilute 10^5 (adjust for measured OD) and then plate 100 ul. You'll likely hit the right place if you did this with 10^4, 10^5, and 10^6 dilutions of an OD 1 culture.


Phase of moon?
uh?

#6 phage434

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Posted 26 August 2012 - 01:17 PM

Phase of moon indicating unknown factors that are affecting your experiment. A joke. I hope.





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