Highly sensitive northern hybridization of rare mRNA using a positively charged nylon membrane
The effect of different RNA fixation methods using Hybond-N and Hybond-N+ nylon membranes in Northern blot experiments was analyzed with human glyceraldehyde-3-phosphate dehydrogenase and human interleukin-3 receptor alpha-subunit as radioactive labeled DNA probes. Highest sensitivity could be achie...
| Veröffentlicht in: | BioTechniques. - 1988. - 16(1994), 6 vom: 30. Juni, Seite 1074-8 |
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| Weitere Verfasser: | , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
1994
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Technical Report Journal Article DNA Probes Membranes, Artificial Nylons RNA, Messenger Receptors, Interleukin-3 RNA 63231-63-0 Glyceraldehyde-3-Phosphate Dehydrogenases |
| Zusammenfassung: | The effect of different RNA fixation methods using Hybond-N and Hybond-N+ nylon membranes in Northern blot experiments was analyzed with human glyceraldehyde-3-phosphate dehydrogenase and human interleukin-3 receptor alpha-subunit as radioactive labeled DNA probes. Highest sensitivity could be achieved by mild alkaline fixation of RNA to positively charged Hybond-N+ nylon membrane. Fixation by UV light resulted in reduced sensitivity in comparison with alkaline treatment. Fixation of RNA to Hybond-N or -N+ membranes by baking for 2 h at 80 degrees C resulted in a strong decrease in sensitivity and should be avoided when using modified nylon membranes in Northern hybridizations |
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| Beschreibung: | Date Completed 06.10.1994 Date Revised 17.04.2006 published: Print Citation Status MEDLINE |
| ISSN: | 0736-6205 |