Abstract
Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes.
| Original language | English |
|---|---|
| Pages (from-to) | 327-333 |
| Number of pages | 7 |
| Journal | Bioconjugate Chemistry |
| Volume | 19 |
| Issue number | 1 |
| DOIs | |
| Publication status | Published - 2008 Jan |
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ASJC Scopus subject areas
- Chemistry(all)
- Organic Chemistry
- Clinical Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
Cite this
Rapid DNA chemical ligation for amplification of RNA and DNA signal. / Abe, Hiroshi; Kondo, Yuko; Jinmei, Hiroshi; Abe, Naoko; Furukawa, Kazuhiro; Uchiyama, Atsushi; Tsuneda, Satoshi; Aikawa, Kyoko; Matsumoto, Isamu; Ito, Yoshihiro.
In: Bioconjugate Chemistry, Vol. 19, No. 1, 01.2008, p. 327-333.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Rapid DNA chemical ligation for amplification of RNA and DNA signal
AU - Abe, Hiroshi
AU - Kondo, Yuko
AU - Jinmei, Hiroshi
AU - Abe, Naoko
AU - Furukawa, Kazuhiro
AU - Uchiyama, Atsushi
AU - Tsuneda, Satoshi
AU - Aikawa, Kyoko
AU - Matsumoto, Isamu
AU - Ito, Yoshihiro
PY - 2008/1
Y1 - 2008/1
N2 - Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes.
AB - Enzymatic ligation methods are useful in the diagnostic detection of DNA sequences. Here, we describe the investigation of nonenzymatic phosphorothioate-iodoacetyl DNA chemical ligation as a method for the detection and identification of RNA and DNA. The specificity of ligation on the DNA target is shown to allow the discrimination of a single point mutation with a drop in the ligation yield of up to 16.1-fold. Although enzymatic ligation has very low activity for RNA targets, this reaction is very efficient for RNA targets. The speed of the chemical ligation with an RNA target achieves a 70% yield in 5 s, which is equal to or better than that of ligase-enzyme-mediated ligation with a DNA target. The reaction also exhibits a significant level of signal amplification under thermal cycling in periods as short as 100-120 min, with the RNA or DNA target acting in a catalytic way to ligate multiple pairs of probes.
UR - http://www.scopus.com/inward/record.url?scp=38949192864&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=38949192864&partnerID=8YFLogxK
U2 - 10.1021/bc700244s
DO - 10.1021/bc700244s
M3 - Article
C2 - 17990846
AN - SCOPUS:38949192864
VL - 19
SP - 327
EP - 333
JO - Bioconjugate Chemistry
JF - Bioconjugate Chemistry
SN - 1043-1802
IS - 1
ER -