| Summary: | 碩士 === 國立中興大學 === 畜牧學系 === 84 === Studies on RAPD and RAMPO Fingerprintsof Holstein Cattle
Abstract Random amplified polymorphic DNA (RAPD) is amplified
by polymerase chain reaction device . A single, short random
oligonucleiotide primer is used to amplified no-specific gene of
genomic DNA. Because that the primer is very short, so it can
anneal at many different loci. If two primers extended at
correct direction in DNA template, random DNA fragment can be
generated. RAPD fingerprints is a dominant genetic marker which
were inherited according to Mendelian law. Random amplified
microsatellite polymorphisms (RAMPO) technique combines RAPD
with microsatellite hybridization to produce several
independence and polymorphic genetic fingerprints per
electrophoretic gel. The RAMPO fingerprints are also inherited
according to the Mendelian law. The purpose of this experiment
was to study on the optimization of RAPD-PCR in Holstein cattle,
and to compare double-primer, one after another additive
different primers, RAPD products digested with restriction
enzymes and RAPD-PCR after template DNA be digested to establish
RAPD fingerprints of different sex or different levels of milk
production, and DNA fingerprint analysis were made with RAMPO
fingerprints to compare the difference of sex or milk
production. The results showed that the optimization of RAPD-
PCR in Holstein cattle where as follows, the concentration of
template DNA : 100 ng/25 ul,the concentration of primer : 0.14
uM, the concentration of polymerase : 0.5 U/25 ul, the
concentration of dNTPs : 400 uM and the concentration of MgCl2 :
1.5 mM. Eighty-seven percentage of random primers of OPAA,OPAO,
OPAV, OPC and OPE sceries used for RAPD-PCR could generate RAPD
products.RAMPO fingerprints in Holstein cattle were
polymorphisms. The RAMPO fingerprints of Holstein cattle showed
a sex-specific band presented in the male only when genomic DNA
was amplified with OPE-01 primer and hybridized with probe
(TG)6.
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