difference between subculture in incubator and shaker - on preparation of cultures for plasmid dna extraction, is there a diff (May/27/2008 )
Hi all,
I'm subculturing some multi-resistant staphylococcus strains for plasmid dna extraction. I'll be using Qiagen plasmid dna extraction kit which requires bacterial strains to be subcultured in Lauria-bertani broth for 12-16 hours overnight with shaking . Unfortunately there was a problem and i couldn't find an available shaker so i subcultured them in a conventional incubator instead. I was supposed to perform the extraction tomorrow. The question is: Is there a significant difference between subcultring in an incubator and in a shaker? Could subculturing without shaking lead to less plasmid propagation for example? I've three options here:
1-If i find considerable growth in the morning, use it and continue extraction with the growth.
2- Throw away the culture and repeat when a shaker is available.
3-Remove the cultures in the morning and put them in a shaker for almost 2 hours in the morning before extraction.
What is your advice? Does anybody have a similar experience?
Thanks in advance
Shaking is very important, bacteria need oxygen to grow.
Just do o/n culture againwhen the shaker is available for optimum bacterial growth and plasmid isolation.
Multiple-resistant staph? Don't ever come nearer to my shaker. 
Just do o/n culture againwhen the shaker is available for optimum bacterial growth and plasmid isolation.
Multiple-resistant staph? Don't ever come nearer to my shaker.
Not sure about the two hour shaking 'cos if it doesn't grow well after O/N culture in incubator, chances are the two hour shaking won't give an big increase.
cellcounter, do cultures get contaminated easily in the shaker? Meaning if one guy has this super-resistant bug, will it actually contaminate the shaker's other cultures?
Just do o/n culture againwhen the shaker is available for optimum bacterial growth and plasmid isolation.
Multiple-resistant staph? Don't ever come nearer to my shaker.
Not sure about the two hour shaking 'cos if it doesn't grow well after O/N culture in incubator, chances are the two hour shaking won't give an big increase.
cellcounter, do cultures get contaminated easily in the shaker? Meaning if one guy has this super-resistant bug, will it actually contaminate the shaker's other cultures?
Well, bugs surely would, it depends upon
1. how secure are the lids,
2. how much culture media
3. in what size flask,
4. how fast and furious it is shaken etc.
Bottomline: there should be NO aerosol formation, especially since the lids are not to be secured tightly to allow aeration.
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