Direct DNA and PNA probe binding to telomeric regions without classical in situ hybridization

Molecular Cytogenetics

Takamitsu A Kato¹  Ian M Cartwright¹  Matthew D Genet¹ 

[1] Department of Environmental and Radiological Health Sciences, Colorado State University, 1618 Campus Delivery, Fort Collins, CO 80523, USA

关键词: Cytogenetics;    FISH;    PNA probes;   

Others  :  1150773
DOI  :  10.1186/1755-8166-6-42

 received in 2013-08-08, accepted in 2013-09-11,  发布年份 2013

【 摘 要 】

Background

Fluorescence in situ Hybridization (FISH) utilizes peptide nucleic acid (PNA) probes to identify specific DNA sequences. Traditional techniques have required the heat denaturing of the DNA in formamide followed by multiple hours at moderated temperatures to allow the probe to hybridize to its specific target. Over the past 30 years, advancements in both protocols and probes have made FISH a more reliable technique for both biological research and medical diagnostics, additionally the protocol has been shortened to several minutes. These PNA probes were designed to target and hybridize to both DNA and RNA, and PNA-protein interactions still remain unclear.

Results

In this study we have shown that a telomeric single stranded specific PNA probe is able to bind to its target without heat denaturing of the DNA and without formamide. We have also identified a centromere specific probe, which was found to bind its target with only incubation with formamide.

Conclusions

Certain PNA probes are able to hybridize with their targets with minimal to no denaturing of the DNA itself. This limited denaturing preserves the chromosome structure and may lead to more effective and specific staining.

【 授权许可】

   
2013 Genet et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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