Impact of RNA isolation protocols on RNA detection by Northern blotting.
Identifieur interne : 001754 ( Main/Exploration ); précédent : 001753; suivant : 001755Impact of RNA isolation protocols on RNA detection by Northern blotting.
Auteurs : Katrin Damm [Allemagne] ; Simone Bach ; Katrin M H. Müller ; Gabriele Klug ; Olga Y. Burenina ; Elena A. Kubareva ; Arnold Grünweller ; Roland K. HartmannSource :
- Methods in molecular biology (Clifton, N.J.) [ 1940-6029 ] ; 2015.
Descripteurs français
- KwdFr :
- MESH :
- isolement et purification : ARN bactérien, Oligonucléotides, Polyribonucléotides.
- normes : Technique de Northern.
- Bacillus subtilis, Phénol, Technique de Northern, Techniques de culture cellulaire.
English descriptors
- KwdEn :
- MESH :
- chemical , isolation & purification : Oligonucleotides, Polyribonucleotides, RNA, Bacterial.
- chemistry : Bacillus subtilis.
- methods : Blotting, Northern.
- standards : Blotting, Northern.
- Cell Culture Techniques, Phenol.
Abstract
We prepared total RNA from the Gram-positive soil bacterium Bacillus subtilis by different RNA extraction procedures to compare their suitability for Northern blot detection of tiny RNAs (~14-mers) or RNAs of intermediate size (100-200 nt) in terms of signal quality, intensity, and reproducibility. Our analysis included two hot phenol methods and two TRIzol extraction procedures. We found that signal intensity/detection sensitivity makes the key difference. Total RNAs prepared by the hot phenol method comprise the length spectrum from tRNAs to large ribosomal RNAs. Larger RNAs are less abundant in TRIzol preparations which instead enrich for RNAs of tRNA size and smaller. Thus, hot phenol methods are the choice for the detection of intermediate-sized and longer RNAs, whereas TRIzol extraction procedures are more suited for the detection of tiny RNAs.
DOI: 10.1007/978-1-4939-2547-6_4
PubMed: 25791588
Affiliations:
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Le document en format XML
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Müller</name></author><author><name sortKey="Klug, Gabriele" sort="Klug, Gabriele" uniqKey="Klug G" first="Gabriele" last="Klug">Gabriele Klug</name></author><author><name sortKey="Burenina, Olga Y" sort="Burenina, Olga Y" uniqKey="Burenina O" first="Olga Y" last="Burenina">Olga Y. Burenina</name></author><author><name sortKey="Kubareva, Elena A" sort="Kubareva, Elena A" uniqKey="Kubareva E" first="Elena A" last="Kubareva">Elena A. Kubareva</name></author><author><name sortKey="Grunweller, Arnold" sort="Grunweller, Arnold" uniqKey="Grunweller A" first="Arnold" last="Grünweller">Arnold Grünweller</name></author><author><name sortKey="Hartmann, Roland K" sort="Hartmann, Roland K" uniqKey="Hartmann R" first="Roland K" last="Hartmann">Roland K. 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Our analysis included two hot phenol methods and two TRIzol extraction procedures. We found that signal intensity/detection sensitivity makes the key difference. Total RNAs prepared by the hot phenol method comprise the length spectrum from tRNAs to large ribosomal RNAs. Larger RNAs are less abundant in TRIzol preparations which instead enrich for RNAs of tRNA size and smaller. Thus, hot phenol methods are the choice for the detection of intermediate-sized and longer RNAs, whereas TRIzol extraction procedures are more suited for the detection of tiny RNAs. </div></front></TEI><affiliations><list><country><li>Allemagne</li></country><region><li>District de Giessen</li><li>Hesse (Land)</li></region><settlement><li>Marbourg</li></settlement></list><tree><noCountry><name sortKey="Bach, Simone" sort="Bach, Simone" uniqKey="Bach S" first="Simone" last="Bach">Simone Bach</name><name sortKey="Burenina, Olga Y" sort="Burenina, Olga Y" uniqKey="Burenina O" first="Olga Y" last="Burenina">Olga Y. Burenina</name><name sortKey="Grunweller, Arnold" sort="Grunweller, Arnold" uniqKey="Grunweller A" first="Arnold" last="Grünweller">Arnold Grünweller</name><name sortKey="Hartmann, Roland K" sort="Hartmann, Roland K" uniqKey="Hartmann R" first="Roland K" last="Hartmann">Roland K. Hartmann</name><name sortKey="Klug, Gabriele" sort="Klug, Gabriele" uniqKey="Klug G" first="Gabriele" last="Klug">Gabriele Klug</name><name sortKey="Kubareva, Elena A" sort="Kubareva, Elena A" uniqKey="Kubareva E" first="Elena A" last="Kubareva">Elena A. Kubareva</name><name sortKey="Muller, Katrin M H" sort="Muller, Katrin M H" uniqKey="Muller K" first="Katrin M H" last="Müller">Katrin M H. Müller</name></noCountry><country name="Allemagne"><region name="Hesse (Land)"><name sortKey="Damm, Katrin" sort="Damm, Katrin" uniqKey="Damm K" first="Katrin" last="Damm">Katrin Damm</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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