14 Polymerase Chain Reaction
219
• Arbitrarily primed PCR (APPCR) or random amplified polymorphic
DNA (RAPD) use short random primer mixes to characterize cloned
fragments of genomic DNA (20, 21).
• Sequence tagged sites (STSs) are genomic Iandmarks at lOOkb distance
(22) as defined by the human genome projects.
• PCR across gene rearrangements is widely used in viral diagnostics,
break point determination in oncogenes (84), and determination of rearrangents in immunoglobulin genes.
• "Footprinting" is used to identify binding regions for DNA-binding proteins (23).
• Microdissection by PCR (24) permits the generation of probes for fluorescence in-situ hybridization of chromosomal segments.
• PCR on single cells (25).
RNA template
PCR can be applied to the study of gene expression. Starting from mRNA
and using reverse transcriptase the first cDNA strand is synthesized, which
then serves as the template for the PCR.
• Anchored PCR (26) or "RACE: rapid amplification of cDNA ends" (27) is
a two-step approach to clone a full-length cDNA (see also "PCR-cloning").
• Multiple oligonucleotide primed amplification of cDNA (MOPAC) (28)
uses degenerate primer mixtures to clone novel genes or cDNAs under
the hypothesis of sequence similarity between gene families and genes
from different species (29).
• mRNA phenotyping is a method for studying the level of expression of
specific transcripts from specific cell types within a heterogeneaus cell
population (30).
• In-situ PCR represents a PCR-based approach to in situ hybridization
(31).
• PCR permits the creation of specific libraries from a sub-population of
cells.
219
• Arbitrarily primed PCR (APPCR) or random amplified polymorphic
DNA (RAPD) use short random primer mixes to characterize cloned
fragments of genomic DNA (20, 21).
• Sequence tagged sites (STSs) are genomic Iandmarks at lOOkb distance
(22) as defined by the human genome projects.
• PCR across gene rearrangements is widely used in viral diagnostics,
break point determination in oncogenes (84), and determination of rearrangents in immunoglobulin genes.
• "Footprinting" is used to identify binding regions for DNA-binding proteins (23).
• Microdissection by PCR (24) permits the generation of probes for fluorescence in-situ hybridization of chromosomal segments.
• PCR on single cells (25).
RNA template
PCR can be applied to the study of gene expression. Starting from mRNA
and using reverse transcriptase the first cDNA strand is synthesized, which
then serves as the template for the PCR.
• Anchored PCR (26) or "RACE: rapid amplification of cDNA ends" (27) is
a two-step approach to clone a full-length cDNA (see also "PCR-cloning").
• Multiple oligonucleotide primed amplification of cDNA (MOPAC) (28)
uses degenerate primer mixtures to clone novel genes or cDNAs under
the hypothesis of sequence similarity between gene families and genes
from different species (29).
• mRNA phenotyping is a method for studying the level of expression of
specific transcripts from specific cell types within a heterogeneaus cell
population (30).
• In-situ PCR represents a PCR-based approach to in situ hybridization
(31).
• PCR permits the creation of specific libraries from a sub-population of
cells.