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FRIEDHELM HILDEBRANDT AND IVA SINGH-SAWHNEY
Applications
Modifications and applications of PCR for specific purposes
PCR is a veryversatile method. A multitude of modifications of the PCR have
been successfully applied for experimental purposes. In this section such
special applieations of PCR are listed, classified according to emphasis
on the use of a specific template, primer, cycling protocol, product detection
method, quantitation of PCR, or polymerase used. Many of these methods
represent approaches to partiallyovercomethe most severelimitation to PCR,
the requirement for specific sequence information flankingthe target region.
Genomic template
• PCR is used in prenatal diagnosis on chorionie villus biopsy specimens,
amniocytes, in vitro fertilized eggs, prefertilization polar bodies, and on
matemal peripheral blood and preimplantation embryos (8) for sex-determination of the fetus.
• PCR is used on degraded genomic DNA ("jumping PCR" (9)), on fossil
DNA (10), and on paraffin embedded samples (11).
• PCR is used in forensie applications on single hair, sperm etc ..
• Inverse PCR (12, 13, 14) is used if sequence information is available only
on one side of the target region. The template is digested with a restrietion enzyme and circularized by ligation. PCR is then performed with 2
primers whose 3' ends point away from each other, thereby amplifying
the sequence of the circle.
• MVR-PCR (15) whieh permits unequivocal identification of an individual through genotyping of an extremely polymorphie marker is mainly
used for forensie applieations.
• Mierosatellite-PCR markers permit genotyping of highly polymorphie
dinucleotide-repeat markers for use in linkage analysis (16).
• Inter-Alu repeat PCR employs primer complementary to repetitive sequences (IRSs) of genomie DNA to characterize overlapping YAC clones
(17).
• Transposon-related PCR strategies (18, 19) use transposons to provide
sequence information after integration of the transposon into !arge
cloned fragments of genomie DNA.
FRIEDHELM HILDEBRANDT AND IVA SINGH-SAWHNEY
Applications
Modifications and applications of PCR for specific purposes
PCR is a veryversatile method. A multitude of modifications of the PCR have
been successfully applied for experimental purposes. In this section such
special applieations of PCR are listed, classified according to emphasis
on the use of a specific template, primer, cycling protocol, product detection
method, quantitation of PCR, or polymerase used. Many of these methods
represent approaches to partiallyovercomethe most severelimitation to PCR,
the requirement for specific sequence information flankingthe target region.
Genomic template
• PCR is used in prenatal diagnosis on chorionie villus biopsy specimens,
amniocytes, in vitro fertilized eggs, prefertilization polar bodies, and on
matemal peripheral blood and preimplantation embryos (8) for sex-determination of the fetus.
• PCR is used on degraded genomic DNA ("jumping PCR" (9)), on fossil
DNA (10), and on paraffin embedded samples (11).
• PCR is used in forensie applications on single hair, sperm etc ..
• Inverse PCR (12, 13, 14) is used if sequence information is available only
on one side of the target region. The template is digested with a restrietion enzyme and circularized by ligation. PCR is then performed with 2
primers whose 3' ends point away from each other, thereby amplifying
the sequence of the circle.
• MVR-PCR (15) whieh permits unequivocal identification of an individual through genotyping of an extremely polymorphie marker is mainly
used for forensie applieations.
• Mierosatellite-PCR markers permit genotyping of highly polymorphie
dinucleotide-repeat markers for use in linkage analysis (16).
• Inter-Alu repeat PCR employs primer complementary to repetitive sequences (IRSs) of genomie DNA to characterize overlapping YAC clones
(17).
• Transposon-related PCR strategies (18, 19) use transposons to provide
sequence information after integration of the transposon into !arge
cloned fragments of genomie DNA.