M13 phage amplification,who canhelp?
Posted 25 May 2001 - 09:00 PM
Posted 25 July 2002 - 07:51 PM
I don't want to insult you by giving to simple of an answer, but did you add e.coli cells to the top agar too so the phage would have cells to infect and replicate in?
Posted 31 July 2002 - 11:55 PM
My my perspective, the problem may be:
- the temperature of your top agar is too hot, i.e. far
>>45 degree and the XL1-blue cell may die!!! (or maybe you've already prepare a stock of top agar and use simply by microwave the top agar then put it into the water and use within a short period of time! so the temperature would probably unfavable to the growth condition of the E.coli.
Maybe you can try using lower temperature top agar. i.e. 45 degree even it's quite readily solidified.