Introduction - Genetic Sex Differentiation in Fish - page 15

Table 6. contd....

Real-time
polymerase chain
reaction
(Real time PCR/RT
PCR)

Higuchi et al.
(1992)

It is also called quantitative real time polymerase
chain reaction (Q-PCR/qPCR/qRT-PCR) or kinetic
polymerase chain reaction (KPCR), amplifi es and
quantifi es a targeted DNA molecule, enables both
detection and quantifi cation of the amplifi ed
DNA as the reaction progresses in real time. It
quantitatively measures starting amounts of
DNA, cDNA, or RNA. Quantitative real-time PCR
has a very high degree of precision, e.g., mRNA
measurement in single cells (Bengtsson et al., 2008).
QRT-PCR methods use fl uorescent dyes, such as
Sybr Green, EvaGreen or fl uorophore-containing
DNA probes, such as TaqMan, to measure the
amount of amplifi ed product in real time.

Rapid Amplifi cation
of cDNA Ends
(RACE PCR)

Sambrook
and Russell
(2001)

This technique is used to obtain the full length
sequence of an RNA transcript found within a
cell. RACE results in the production of a cDNA
copy of the RNA sequence of interest and is
produced through reverse transcription, followed
by PCR amplifi cation of the cDNA copies. RACE
can provide the sequence of an RNA transcript
from a small known sequence within the transcript
to the 5' end (5' RACE-PCR) or 3' end (3' RACE-
PCR) of the RNA. This technique is also called as
one-sided PCR or anchored PCR .

Microarray Augenlicht

and Kobrin
(1982)

This technology evolved from Southern
blotting, where fragmented DNA is attached to
a substrate and then probed with a known gene or
fragment. The fi rst reported use of this approach is
the analysis of 378 arrayed bacterial colonies each
harboring a gene. Subsequently this technology
has been extensively evolved and current versions
can analyze more than a million genomic regions
in a single experiment. Three major types of arrays
are: expression arrays, copy number arrays and
SNP arrays. The probes are attached on glass
slides and hybridized with the RNA/DNA sample
of interest and thus the expression status of the
genomic regions spotted on the microarray chips
are inferred.

Trangenesis Palmitter et

al. (1982)

Successful production of transgenic mouse by
introducing metalothionein-human growth
hormone fusion gene ( hGH ) into mouse egg
resulting in dramatic increase in growth. The
restricted scope of cytoplasmic introduction of
transgene is one of hurdles in transgenic fi sh
production. The use of triploid fi sh eggs avoids the
propagation of escaped transgenics (Pandian and
Marian, 1994).