scholarly journals Use of multilocus methylation-specific single nucleotide primer extension (MS-SNuPE) technology in diagnostic testing for human imprinted loci

Epigenetics ◽  
2012 ◽  
Vol 7 (5) ◽  
pp. 473-481 ◽  
Author(s):  
Matthias Begemann ◽  
Isabelle Leisten ◽  
Lukas Soellner ◽  
Klaus Zerres ◽  
Thomas Eggermann ◽  
...  
Keyword(s):  
Diagnostic Testing ◽  
Primer Extension ◽  
Single Nucleotide ◽  
Single Nucleotide Primer Extension

scholarly journals Sensitive quantitative assay for point mutations in the rat H-ras gene based on single nucleotide primer extension

2010 ◽  
Vol 1 (4) ◽  
pp. 657-661 ◽  
Author(s):  
YOSHIHISA YANO ◽  
TOMOHIRO YANO ◽  
ANNA KINOSHITA ◽  
AI MATOBA ◽  
TADAYOSHI HASUMA ◽  
...  
Keyword(s):  
Point Mutations ◽  
Primer Extension ◽  
Quantitative Assay ◽  
Ras Gene ◽  
Single Nucleotide ◽  
Single Nucleotide Primer Extension

scholarly journals Mouse endogenous X-linked genes do not show lineage-specific delayed inactivation during development

1995 ◽  
Vol 65 (3) ◽  
pp. 223-227 ◽  
Author(s):  
Jeanne M. Lebon ◽  
Patrick P. L. Tam ◽  
Judith Singer-Sam ◽  
Arthur D. Riggs ◽  
Seong-Seng Tan
Keyword(s):  
X Chromosome ◽  
Female Mouse ◽  
X Chromosome Inactivation ◽  
Mouse Embryos ◽  
Primer Extension ◽  
Rt Pcr ◽  
Single Nucleotide ◽  
Allele Specific ◽  
Endogenous Genes ◽  
Single Nucleotide Primer Extension

SummaryX chromosome inactivation (XCI) has been assumed to be complete in all cells of female mouse embryos at about 6 d post coitum (dpc). However, a recent study on β-galactosidase expression of an X-linkedlacZtransgene suggests that XCI is probably not complete several days after this time in some lineages. To help resolve this issue, we analysed XCI in embryos which carry the T(X;16)16H (Searle's) translocation and are heterozygous at the X-linkedHprtandPgk-1genes. The quantitative RT-PCR single nucleotide primer extension (SNuPE) assay was used to measureHprtandPgk-1allele-specific transcripts in embryos 9·5 dpc. No transcripts from the normal X chromosome were found in any of the tissues tested, indicating that inactivation was complete for these endogenous genes.

scholarly journals Evaluation of Single-Nucleotide Primer Extension for Detection and Typing of Phylogenetic Markers Used for Investigation of Microbial Communities

2009 ◽  
Vol 75 (9) ◽  
pp. 2850-2860 ◽  
Author(s):  
Marcell Nikolausz ◽  
Antonis Chatzinotas ◽  
András Táncsics ◽  
Gwenaël Imfeld ◽  
Matthias Kästner
Keyword(s):  
16S Rrna ◽  
Dna Sequences ◽  
Simultaneous Detection ◽  
Primer Extension ◽  
Detection Sensitivity ◽  
High Signal ◽  
Rrna Gene ◽  
Single Nucleotide ◽  
Specificity And Sensitivity ◽  
Single Nucleotide Primer Extension

ABSTRACT Single-nucleotide primer extension (SNuPE) is an emerging tool for parallel detection of DNA sequences of different target microorganisms. The specificity and sensitivity of the SNuPE method were assessed by performing single and multiplex reactions using defined template mixtures of 16S rRNA gene PCR products obtained from pure bacterial cultures. The mismatch discrimination potential of primer extension was investigated by introducing different single and multiple primer-target mismatches. The type and position of the mismatch had significant effects on the specificity of the assay. While a 3′-terminal mismatch has a considerable effect on the fidelity of the extension reaction, the internal mismatches influenced hybridization mostly by destabilizing the hybrid duplex. Thus, carefully choosing primer-mismatch positions should result in a high signal-to-noise ratio and prevent any nonspecific extension. Cyclic fluorescent labeling of the hybridized primers via extension also resulted in a significant increase in the detection sensitivity of the PCR. In multiplex reactions, the signal ratios detected after specific primer extension correlated with the original template ratios. In addition, reverse-transcribed 16S rRNA was successfully used as a nonamplified template to prove the applicability of SNuPE in a PCR-independent manner. In conclusion, this study demonstrates the great potential of SNuPE for simultaneous detection and typing of various nucleic acid sequences from both environmental and engineered samples.

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