PMID- 10839126
OWN - NLM
STAT- MEDLINE
DCOM- 20000920
LR  - 20190831
IS  - 1387-2273 (Print)
IS  - 1387-2273 (Linking)
VI  - 741
IP  - 1
DP  - 2000 Apr 28
TI  - Monitoring hybridization during polymerase chain reaction.
PG  - 3-13
AB  - The polymerase chain reaction (PCR) is usually analyzed by gel electrophoresis 
      for size separation of PCR products. Additional separation techniques, such as 
      single-stranded conformational polymorphism (SSCP), denaturing gradient gel 
      electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE) and 
      denaturing high-performance liquid chromatography (DHPLC), can also be used to 
      scan for sequence alterations. These techniques are all based on the effect of 
      PCR product hybridization on mobility. Hybridization can also be monitored with 
      fluorescence during PCR without chromatographic or electrophoretic separation. 
      Continuous monitoring of PCR allows the detection, quantification and sequence 
      specificity of PCR products to be assessed, often without any need for further 
      analysis. In such a closed system, PCR quantification with sensitivity to the 
      single copy level can be achieved using either double-stranded DNA binding dyes 
      or fluorescently labeled allele-specific oligonucleotide (ASO) probes. Melting 
      curve analysis with ASO probes can be used to genotype various alleles, including 
      single base alterations. The integration of rapid cycle PCR and ASO probes in an 
      automated system greatly facilitates research and clinical applications of 
      nucleic acid analysis in genetics, oncology, and infectious disease.
FAU - de Silva, D
AU  - de Silva D
AD  - Idaho Technology Inc., Salt Lake City, UT 84108, USA.
FAU - Wittwer, C T
AU  - Wittwer CT
LA  - eng
PT  - Journal Article
PT  - Review
PL  - Netherlands
TA  - J Chromatogr B Biomed Sci Appl
JT  - Journal of chromatography. B, Biomedical sciences and applications
JID - 9714109
SB  - IM
MH  - *Nucleic Acid Hybridization
MH  - Polymerase Chain Reaction/*methods
RF  - 42
EDAT- 2000/06/06 09:00
MHDA- 2000/09/23 11:01
CRDT- 2000/06/06 09:00
PHST- 2000/06/06 09:00 [pubmed]
PHST- 2000/09/23 11:01 [medline]
PHST- 2000/06/06 09:00 [entrez]
AID - 10.1016/s0378-4347(00)00055-4 [doi]
PST - ppublish
SO  - J Chromatogr B Biomed Sci Appl. 2000 Apr 28;741(1):3-13. doi: 
      10.1016/s0378-4347(00)00055-4.