In addition, because silver staining is a relatively insensitive
method of DNA detection, large amounts of PCR product
were necessary for analysis and resolution tended to be low.
The limitations of the existing methodology led us to develop
an improved version of RISA, which we refer to as automated
RISA (ARISA). This method is similar to the recently published
terminal restriction fragment length polymorphism and
length heterogeneity analysis by PCR community analysis techniques
(13, 24). In the automated approach, the initial steps of
DNA extraction and PCR amplification are the same as in
RISA, except that PCR is conducted with a fluorescencetagged
oligonucleotide primer. The electrophoretic step is
subsequently performed with an automated system, which provides
laser detection of fluorescent DNA fragments. ARISAPCR
may generate DNA fragments up to 1,400 bp in length (6).
Discrimination of these larger size fragments represented a new
application for the capillary electrophoresis system employed.
In addition, because silver staining is a relatively insensitive
method of DNA detection, large amounts of PCR product
were necessary for analysis and resolution tended to be low.
The limitations of the existing methodology led us to develop
an improved version of RISA, which we refer to as automated
RISA (ARISA). This method is similar to the recently published
terminal restriction fragment length polymorphism and
length heterogeneity analysis by PCR community analysis techniques
(13, 24). In the automated approach, the initial steps of
DNA extraction and PCR amplification are the same as in
RISA, except that PCR is conducted with a fluorescencetagged
oligonucleotide primer. The electrophoretic step is
subsequently performed with an automated system, which provides
laser detection of fluorescent DNA fragments. ARISAPCR
may generate DNA fragments up to 1,400 bp in length (6).
Discrimination of these larger size fragments represented a new
application for the capillary electrophoresis system employed.
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