A simplified CsCL protocol for lambda DNA purification: no enzymatic treatment/one phenol extraction

Santelli, Roberto Vicente; Navarro-Cattapan, Luci Deise

doi:10.1590/s1415-47572000000100011

Cited by 4 publications

(3 citation statements)

References 2 publications

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“…The protocol followed was a modification of CsCl lambda DNA purification described in Santelli and Navarro-Cattapan (2000) and in Andrade et al (2005).…”

Section: Rhynchosciara Americana Genomic Phage Library Screeningmentioning

confidence: 99%

Molecular characterization of a putative heat shock protein cognate gene in Rhynchosciara americana

et al. 2009

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An hsc70 homologue gene (Rahsc70) of the diptera Rhynchosciara americana was isolated and characterized. We were able to determine the mRNA sequence from an EST of salivary gland cDNA library, and a Rahsc70 cDNA cassette was used as a probe to isolate the genomic region from a genomic library. The mRNA expression of this gene parallels the 2B puff expansion, suggesting its involvement in protein processing, since this larval period corresponds to a high synthetic activity period. During heat shock stress conditions, hsc70 expression decreased. In situ hybridization of polytene chromosomes showed that the Rahsc70 gene is located near the C3 DNA puff. The cellular localization of Hsc70 protein showed this protein in the cytoplasm and in the nucleus.

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“…The protocol followed was a modification of CsCl lambda DNA purification described in Santelli and Navarro-Cattapan (2000) and in Andrade et al (2005).…”

Section: Rhynchosciara Americana Genomic Phage Library Screeningmentioning

confidence: 99%

Molecular characterization of a putative heat shock protein cognate gene in Rhynchosciara americana

et al. 2009

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“…Washes were done four times at 65-C with 2Â SSC, 0.1% SDS and twice with 0.1Â SSC, 0.1% SDS and exposed to a PhosphorImager system (Storm, Molecular Dynamics). After the first screening, the protocol for isolation of the positive recombinant was as described by Andrade et al (2005) and the lambda DNA purification was done following the protocol described by Santelli & Navarro-Cattapan (2000).…”

Section: Library Screeningmentioning

confidence: 99%

Molecular characterization of a retrotransposon in the Rhynchosciara americana genome and its association with telomere

et al. 2008

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Non-LTR retrotransposons, also known as long interspersed nuclear elements (LINEs), are transposable elements that encode a reverse transcriptase and insert into genomic locations via RNA intermediates. The sequence analysis of a cDNA library constructed from mRNA of the salivary glands of R. americana showed the presence of putative class I elements. The cDNA clone with homology to a reverse transcriptase was the starting point for the present study. Genomic phage was isolated and sequenced and the molecular structure of the element was characterized as being a non-LTR retrotransposable element. Southern blot analysis indicated that this transposable element is represented by repeat sequences in the genome of R. americana. Chromosome tips were consistently positive when this element was used as probe in in-situ hybridization. Real-time RT-PCR showed that this retrotransposon is transcribed at different periods of larval development. Most interesting, the silencing of this retrotransposon in R. americana by RNA interference resulted in reduced transcript levels and in accelerated larval development.

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“…This modification, requiring less steps and fewer solutions to prepare, produces a DNA suitable for further manipulation like digestion, ligation, labelling, subcloning, etc. (Santelli and Navarro-Cattapan, 2000). Now we are proposing a shortcut for the Benton & Davis technique (Benton and Davis, 1977).…”

mentioning

confidence: 99%

A shortcut in phage screening technique

et al. 2005

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A simplified CsCL protocol for lambda DNA purification: no enzymatic treatment/one phenol extraction

Cited by 4 publications

References 2 publications

Molecular characterization of a putative heat shock protein cognate gene in Rhynchosciara americana

Molecular characterization of a putative heat shock protein cognate gene in Rhynchosciara americana

Molecular characterization of a retrotransposon in the Rhynchosciara americana genome and its association with telomere

A shortcut in phage screening technique

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