SEA-PHAGES | Clumping in liquid cultures

Link to this post \| posted 25 Aug, 2016 20:56
welkin	Hello All, Some faculty are reporting clumping in their Gordonia liquid cultures. Here are some ways to get rid of clumps. Ways to get rid of clumps: You can use Tween, like smeg in starter cultures, and then transfer to larger ones. You can start with a small liquid culture and then use it to innoculate a larger one. Using a higher OD culture can help, because you can let any initial clumps settle to the bottom and just innoculate using cells from the top. Make sure you don't grab a big chunky colony (or clump) and use it as part of your innoculum. Clumping is actually so bad in some other Gordonias that they have to be sonicated prior to plating so we can get decent lawns. -Welkin

Link to this post | posted 25 Aug, 2016 20:56

Hello All,
Some faculty are reporting clumping in their Gordonia liquid cultures. Here are some ways to get rid of clumps.

Ways to get rid of clumps:
You can use Tween, like smeg in starter cultures, and then transfer to larger ones.
You can start with a small liquid culture and then use it to innoculate a larger one. Using a higher OD culture can help, because you can let any initial clumps settle to the bottom and just innoculate using cells from the top.

Make sure you don't grab a big chunky colony (or clump) and use it as part of your innoculum.

Clumping is actually so bad in some other Gordonias that they have to be sonicated prior to plating so we can get decent lawns.

-Welkin

Link to this post \| posted 21 Oct, 2018 12:40
smolloy123	Hi All, We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot! Best, Sally

Link to this post | posted 21 Oct, 2018 12:40

smolloy123

Hi All,
We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot!
Best,
Sally

Link to this post \| posted 22 Oct, 2018 13:13
viknesh	smolloy123 Hi All, We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot! Best, Sally That's a really neat trick, Sally. Do you know if it works for M. smeg too? I'll definitely give it a try and report back.

Link to this post | posted 22 Oct, 2018 13:13

viknesh

smolloy123
Hi All,
We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot!
Best,
Sally

That's a really neat trick, Sally. Do you know if it works for M. smeg too? I'll definitely give it a try and report back.

Link to this post \| posted 24 Oct, 2018 15:35
smolloy123	Viknesh Sivanathan smolloy123 Hi All, We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot! Best, Sally That's a really neat trick, Sally. Do you know if it works for M. smeg too? I'll definitely give it a try and report back. Hi Vic, It works with M. smegmatis and fairly well with some of the clumpier species of Mycobacterium. The really clumpy ones sometimes I still need to grow with tween first. Cheers!

Link to this post | posted 24 Oct, 2018 15:35

smolloy123

Viknesh Sivanathan
smolloy123
Hi All,
We have been growing G. terrae in PYCa broth for four days with sterile glass beads in baffled flasks. They plate like buttah! I take the glass 3-mm beads we use for archiving (a bottle is super cheap), sterilize then in culture tubes so that I can easily pour them into flasks, and I do not use Tween ( I am too lazy to take time to sub-culture). After four days of growth, the cultures are smooth and produce perfect lawns! I wash beads and re-autoclave for re-use. Just don't get these re-used beads mixed up with the beads you would use for archiving as those should not be recycled beads. I meant to post this over the summer when I discovered this but forgot!
Best,
Sally

That's a really neat trick, Sally. Do you know if it works for M. smeg too? I'll definitely give it a try and report back.

Hi Vic,
It works with M. smegmatis and fairly well with some of the clumpier species of Mycobacterium. The really clumpy ones sometimes I still need to grow with tween first.
Cheers!

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Clumping in liquid cultures