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Combining sequence-specific probes and DNA binding dyes in real-time PCR for specific nucleic acid quantification and melting curve analysis

Lind, K ; Stålberg, Anders LU ; Zoric, N and Kubista, M (2006) In BioTechniques 40(3). p.315-319
Abstract
Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has tire advantage that it only detects the targeted product, while the nonspecific dye has the advantage that melting curve analysis can be performed after completed amplifcation, which reveals what kind of products have been formed. Here we present a new strategy based on combining a sequence-specific probe and a nonspecific dye, BOXTO, in the swine reaction, to take the advantage of both chemistries. We show that BOXTO can be used together with both TaqMan((R)) probes and locked nucleic acid (LNA) probes without... (More)
Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has tire advantage that it only detects the targeted product, while the nonspecific dye has the advantage that melting curve analysis can be performed after completed amplifcation, which reveals what kind of products have been formed. Here we present a new strategy based on combining a sequence-specific probe and a nonspecific dye, BOXTO, in the swine reaction, to take the advantage of both chemistries. We show that BOXTO can be used together with both TaqMan((R)) probes and locked nucleic acid (LNA) probes without interfering with the PCR. The probe signal reflect formation of target product, while melting curve analysis of the BOXTO signal reveals primer-dinter formation and the presence of any other anomalous products. (Less)
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
BioTechniques
volume
40
issue
3
pages
315 - 319
publisher
Informa Healthcare
external identifiers
  • pmid:16568820
  • wos:000236210400009
  • scopus:33646005816
ISSN
0736-6205
DOI
10.2144/000112101
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Stem Cell and Pancreas Developmental Biology (013212044)
id
953c18c4-8625-4a66-8061-029a33d1063a (old id 908750)
alternative location
http://www.biotechniques.com/default.asp?page=article_archive&subsection=article_display&year=2006&issue=3/1/2006&id=112101
date added to LUP
2016-04-01 16:57:46
date last changed
2021-06-30 01:05:31
@article{953c18c4-8625-4a66-8061-029a33d1063a,
  abstract     = {Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has tire advantage that it only detects the targeted product, while the nonspecific dye has the advantage that melting curve analysis can be performed after completed amplifcation, which reveals what kind of products have been formed. Here we present a new strategy based on combining a sequence-specific probe and a nonspecific dye, BOXTO, in the swine reaction, to take the advantage of both chemistries. We show that BOXTO can be used together with both TaqMan((R)) probes and locked nucleic acid (LNA) probes without interfering with the PCR. The probe signal reflect formation of target product, while melting curve analysis of the BOXTO signal reveals primer-dinter formation and the presence of any other anomalous products.},
  author       = {Lind, K and Stålberg, Anders and Zoric, N and Kubista, M},
  issn         = {0736-6205},
  language     = {eng},
  number       = {3},
  pages        = {315--319},
  publisher    = {Informa Healthcare},
  series       = {BioTechniques},
  title        = {Combining sequence-specific probes and DNA binding dyes in real-time PCR for specific nucleic acid quantification and melting curve analysis},
  url          = {http://dx.doi.org/10.2144/000112101},
  doi          = {10.2144/000112101},
  volume       = {40},
  year         = {2006},
}