A heteroduplex-preferential Tm depressor for the specificity-enhanced DNA polymerase chain reactions
Analytical Biochemistry Volume 337 Issue 1
Page 154-160
published_at 2005-02-01
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| File | |
| Title ( eng ) |
A heteroduplex-preferential Tm depressor for the specificity-enhanced DNA polymerase chain reactions
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| Creator |
Kinoshita-Kikuta Emiko
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| Source Title |
Analytical Biochemistry
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| Volume | 337 |
| Issue | 1 |
| Start Page | 154 |
| End Page | 160 |
| Abstract |
A macrocyclic tetraamine zinc(II) complex appended with two quinoline groups, Zn2+.1,7-bis(4-quinolylmethyl)-1,4,7,10-tetraazacyclododecane (Zn2+.Q2-cyclen), was successfully used as a novel additive to suppress non-specific products in DNA polymerase chain reaction (PCR). In the presence of Zn2+.Q2-cyclen, the Tm drop of 20-bp heteroduplexes containing a non-complementary base pair was greater than that of the corresponding homoduplex (i.e., primer DNA). Here, we applied such preferential DNA melting to a specificity-enhanced PCR using micromolar concentrations of Zn2+.Q2-cyclen. We demonstrated the selective amplification of target DNA fragments (i.e., the human heart sodium channel Nav1.5 gene) from genomic DNA or a cDNA library. The optimum condition for the specificity-enhanced PCR could be determined in the concentration range of 1.50 μM of Zn2+.Q2-cyclen.
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| Keywords |
Macrocyclic tetraamine
Zinc(II) complex
Tm depressor
Specificity-enhanced PCR
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| NDC |
Biology [ 460 ]
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| Language |
eng
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| Resource Type | journal article |
| Publisher |
Elsevier
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| Date of Issued | 2005-02-01 |
| Rights |
Copyright (c) 2004 Elsevier Ltd.
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| Publish Type | Author’s Original |
| Access Rights | open access |
| Source Identifier |
[ISSN] 0003-2697
[DOI] 10.1016/j.ab.2004.10.009
[NCID] AA00524867
[PMID] 15649389
[DOI] http://dx.doi.org/10.1016/j.ab.2004.10.009
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