Please help with MS-2 assay - (Nov/05/2011 )
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.
olu on Sun Nov 6 00:16:11 2011 said:
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-online.org/biology-forums/posts/28248.html