Zinc(II)-cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/Thy-rich DNA fragments
Analytical Biochemistry Volume 408 Issue 2
Page 348-350
published_at 2011-01-15
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| File | |
| Title ( eng ) |
Zinc(II)-cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/Thy-rich DNA fragments
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| Creator |
Kinoshita-Kikuta Emiko
Harada Naoko
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| Source Title |
Analytical Biochemistry
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| Volume | 408 |
| Issue | 2 |
| Start Page | 348 |
| End Page | 350 |
| Abstract |
We describe an improved gel-based method with an additive Zn2+-cyclen complex (cyclen, 1,4,7,10-tetraazacyclododecane), Zn2+-cyclen-PAGE, for mutation detection in DNA fragments by PCR that contain more than 65% Ade/Thy bases and fewer than 100 base pairs (bp). Existing techniques have a problem in analyzing such short Ade/Thy-rich fragments because the duplexes are disrupted and are not detectable due to binding of Zn2+-cyclen to Thy bases. In this strategy using a PCR primer with a Gua/Cyt-lined sequence attached at its 5'-end, we successfully detected a mutation in an 86-bp Ade/Thy-rich region of the BRCA1 gene from formalin-fixed paraffin-embedded breast cancer-tissue sections.
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| Keywords |
Zn2+-cyclen
PAGE
SNP
Mutation
FFPE
BRCA1
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| NDC |
Medical sciences [ 490 ]
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| Language |
eng
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| Resource Type | journal article |
| Publisher |
Academic Press Inc Elsevier Science
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| Date of Issued | 2011-01-15 |
| Rights |
Copyright (c) 2010 Elsevier Inc
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| Publish Type | Author’s Original |
| Access Rights | open access |
| Source Identifier |
[ISSN] 0003-2697
[DOI] 10.1016/j.ab.2010.09.003
[NCID] AA00524867
[DOI] http://dx.doi.org/10.1016/j.ab.2010.09.003
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