Guoliang Qian:
Phi X174 is a bacteriophage of Escherichia coli. There is a large amount of
literature on the growth and propagation of this bacteriophage. In brief,
the bacteriophage reproduces as a double-stranded molecule inside the cell,
then produces single-stranded DNA for encapsulation. The bacteriophage
contains single-stranded DNA and is extruded from the cell without lysing
the cells. Thus, a large amount of phage can be produced. It is very
useful in a molecular biological sense, in that you can cut the
double-stranded phage with restriction enzyme(s), add your favorite gene
into the mix, add ligase and then transform E. coli cells with the mixture.
Bacteriophage particles with single-stranded DNA will be produced, if your
ligation and transformation conditions were correct. The usefulness in
molecular biology comes from the ablity to obtain a large amount of a
single-strand of the recombinant DNA of interest. In planning your
experiments, you MUST determine which strand of your DNA is of interest to
you. This will determine in which orientation you want to clone your
favorite gene (the bacteriophage only replicates one strand and all of the
bacteriophage produced contain the same strand of DNA). If you are
interested in using the bacteriophage in molecular biology, then check out
any of the cloning manuals that are available in the library. If you are
using this bacteriophage to study the basic biology of the phage or of E.
coli, then you'll have a wealth of information in your library.
Scott Jamison
Guoliang Qian wrote in message ...
>Dear,
>>I just got some double strand super coiled PhiX 174DNA. As it is very
>important for my research and I have to use it very often, I need to make a
>large amout of it. Since I am not familiar with this subject, can anyone be
>kind enough to tell me the protocol or where I can find the necessary
>information?
>>Thank you very very much
>>>
