General method for amplifying regions of very high G + C content

Dutton, Charyl M.; Paynton, Christine; Sommer, Steve S.

doi:10.1093/nar/21.12.2953

Cited by 63 publications

(41 citation statements)

References 7 publications

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“…Previously, utilization of cosolvents, formamide (2) or glycerol (3) in the PCR was reported to significantly enhance specificity of the amplification. Most recently, a general method using native but not recombinant Pfu DNA polymerase was recommended to amplify regions of very high GC content (4). In the present study, we show that template denaturation with NaOH was required for PCR amplification of a highly GC-rich template.…”

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confidence: 58%

PCR amplification of highly GC-rich DNA template after denaturation by NaOH

Agarwal

Perl

1993

Nucl Acids Res

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confidence: 58%

PCR amplification of highly GC-rich DNA template after denaturation by NaOH

Agarwal

Perl

1993

Nucl Acids Res

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“…These conflicting results may be caused by the biases of both PCR methods and culture-based methods. The guanineplus-cytosine (G + C) content of template DNA has been reported to influence gene amplification by PCR [7,23]. Reysenbach et al (23) found that low G + C rDNA was preferentially amplified from a mixture of low and high G + C rDNA.…”

Section: MD Shawkey Et Al: Microbial Diversity Of Bird Feathersmentioning

confidence: 99%

Microbial Diversity of Wild Bird Feathers Revealed throughCulture-Based and Culture-Independent Techniques

et al. 2005

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Despite recent interest in the interactions between birds and environmental microbes, the identities of the bacteria that inhabit the feathers of wild birds remain largely unknown. We used culture-based and culture-independent surveys of the feathers of eastern bluebirds (Sialis sialis) to examine bacterial flora. When used to analyze feathers taken from the same birds, the two survey techniques produced different results. Species of the poorly defined genus Pseudomonas were most common in the molecular survey, whereas species of the genus Bacillus were predominant in the culture-based survey. This difference may have been caused by biases in both the culture and polymerase chain reaction techniques that we used. The pooled results from both techniques indicate that the overall community is diverse and composed largely of members of the Firmicutes and b-and c-subdivisions of the Proteobacteria. For the most part, bacterial sequences isolated from birds were closely related to sequences of soil-borne and water-borne bacteria in the GenBank database, suggesting that birds may have acquired many of these bacteria from the environment. However, the metabolic properties and optimal growth requirements of several isolates suggest that some of the bacteria may have a specialized association with feathers.

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“…The next 30 cycles were carried out with an annealing temperature of 658C instead of 438C. This method was based on a protocol described previously for PCR ampli®cation of very high GC rich sequences (Dutton et al, 1993).…”

Section: Construction Of C-jun 5' Utr Mutantsmentioning

confidence: 99%