Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo

bacterial culture not pelleting


  • Please log in to reply
4 replies to this topic

#1 neuron

neuron

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 230 posts
6
Neutral

Posted 06 February 2018 - 02:48 AM

Hi,

 

There is really strange thing happening in our lab. We are growing S.aureus in minimal media, and when we try to pellet down the cells, we are not able to. Cells sediments in the bottom with spin, however as soon as you pour the supernatant, they get disrupted. We tried taking out sup with pipette also, but there also it gets disrupted. We are using polypropylene bottles. And this is for 1 liter or more than that.

 

Anybody faced this problem?

 

Please suggest guys, whats wrong here?

 

Thanks



#2 neuron

neuron

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 230 posts
6
Neutral

Posted 06 February 2018 - 09:40 PM

nobody faced this problemsad.png



#3 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,507 posts
542
Excellent

Posted 07 February 2018 - 08:28 AM

I've never worked with S. aureus, other than as a sample in undergrad labs.

 

Have you tried plating it out? If so, has it changed its behavior? It could be that it has acquired a capsule or something like that, that is preventing pelleting firmly. 

 

You could also try pelleting at higher RCF and see if that helps.



#4 neuron

neuron

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 230 posts
6
Neutral

Posted 07 February 2018 - 09:23 PM

Hey Bob, Thanks a lot for giving some insightssmile.png

 

Yeah, we have planned to put the culture in plates to see the behavior of cells. One of my friend suggested that if the cells are forming biofilm, it will not pellet down, to rule out that possibility also, we will be plating the cells.

 

I checked on google that poly carbonate bottles are poor in chemical resistant. And with CaCl2, no information is available if its compatible. The bottles that we are using are pp, so ruled out that also.



#5 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 6,507 posts
542
Excellent

Posted 08 February 2018 - 08:16 AM

polypropylene and polycarbonate should both be fine for pelleting cells. polycarbonate tends to be a little more slippery so cells detach easier, but only slightly. Both should be fine with calcium chloride.






Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.