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Please help with MS-2 assay

E.coli host antibiotics MS-2

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#1 olu

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Posted 05 November 2011 - 04:16 PM

Please, what is the effects of overdosing the E.coli host with antibiotics? Please, I need explanation.
Secondly, Please is there anyone that can help me out with MS-2 assay. I have being having problem with the assay for some months now.

Here is my steps

1. I make a overnite culture of E.coli with 2 ml of antibiotics
2. I make a log phase culture of E.coli host with 2 ml antibiotics (ampicilin and streptomycin).
3. I make a serial dilution of MS-2 in PBS (pH 7.4-7.6) upto 1E-12 dilution
4. Autoclave the 1 % agar at 121 C for 15 mins and allow to cool down to 42-44 C
5. I added 1 ml of E.coli (log phase) per 100 ml of agar. Mix well and transfer ~30 ml of agar to the diluted PBS containing MS-2.
6. Incubate for 37 C overnight inside water jacket incubator that is calibrate at flow rate of 1 L/s.

Please kindly help me with advice.

#2 pito

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Posted 15 November 2011 - 02:20 PM

Please, what is the effects of overdosing the E.coli host with antibiotics? Please, I need explanation.
Secondly, Please is there anyone that can help me out with MS-2 assay. I have being having problem with the assay for some months now.

Here is my steps

1. I make a overnite culture of E.coli with 2 ml of antibiotics
2. I make a log phase culture of E.coli host with 2 ml antibiotics (ampicilin and streptomycin).
3. I make a serial dilution of MS-2 in PBS (pH 7.4-7.6) upto 1E-12 dilution
4. Autoclave the 1 % agar at 121 C for 15 mins and allow to cool down to 42-44 C
5. I added 1 ml of E.coli (log phase) per 100 ml of agar. Mix well and transfer ~30 ml of agar to the diluted PBS containing MS-2.
6. Incubate for 37 C overnight inside water jacket incubator that is calibrate at flow rate of 1 L/s.

Please kindly help me with advice.


Ok, but what are the exact problems?
It seems ok what you do.

Also check this:
http://www.protocol-...osts/28248.html

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






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