Does anyone have experience with LONG PCR?

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Thu Sep 15 13:02:00 EST 1994


 In article <kamin_johnson-0909941524330001 at bongo.cellbio.duke.edu>
 kamin_johnson at cellbio.duke.edu (kamin johnson) writes:

>> We are attempting to amplify a 16 kb fragment and would like to hear
>> about other's experience with using technique, buffer recipes,
>> protocols, and kits tried.

> In a recent issue of TIBS (I think, one of the Trends... issues anyway)
> there was a discussion of the "long and accurate" PCR technique.  The
> point was that the technique seems to require one to follow the protocol
> of Barnes rigidly.  The protocol was given in the issue.

You should probably read these, and then come back with more specifics:

@article{Hengen1994Augtibs,
author = "P. N. Hengen",
title = "Methods and Reagents - Long and accurate PCR",
journal = "Trends in Biochemical Sciences",
volume = "19",
number = "8",
pages = "341-342",
month = "August",
year = "1994"}

@article{Barnes1994,
author = "W. M. Barnes",
title = "{PCR} amplification of up to 35-kb {DNA} with high
fidelity and high yield from $\lambda$ bacteriophage templates",
journal = "Proc. Natl. Acad. Sci. USA",
volume = "91",
pages = "2216-2220",
month = "March",
year = "1994"}

@article{Cheng.Barnes1994,
author = "S. Cheng
     and C. Fockler
     and W. M. Barnes
     and R. Higuchi",
title = "Effective amplification of long targets
from cloned inserts and human genomic DNA",
journal = "Proc. Natl. Acad. Sci. USA",
volume = "91",
number = "12",
pages = "5695-5699",
month = "June",
year = "1994"}

*******************************************************************************
* Paul N. Hengen, Ph.D.                           /--------------------------/*
* National Cancer Institute                       |Internet: pnh at ncifcrf.gov |*
* Laboratory of Mathematical Biology              |   Phone: (301) 846-5581  |*
* Frederick Cancer Research and Development Center|     FAX: (301) 846-5598  |*
* Frederick, Maryland 21702-1201 USA              /--------------------------/*
* - -  Methods FAQ list -> http://fconvx.ncifcrf.gov:2001/~pnh/info.html  - - *
*******************************************************************************



More information about the Methods mailing list