Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA
We have developed a simple and sensitive method for the rapid quantitation of mRNA from cell cultures and small tissue samples. The method combines the high sensitivity and specificity of the ribonuclease protection assay with simple handling and rapid execution of dot blotting. The use of digoxygen...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
1997
|
| In: |
BioTechniques
Year: 1997, Volume: 22, Issue: 3, Pages: 500-505 |
| ISSN: | 1940-9818 |
| DOI: | 10.2144/97223st05 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.2144/97223st05
|
| Author Notes: | Jinghai Zhan, H. Dariush Fahimi, Alfred Völkl |
MARC
| LEADER | 00000caa a2200000 c 4500 | ||
|---|---|---|---|
| 001 | 1914059867 | ||
| 003 | DE-627 | ||
| 005 | 20250716221533.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 250109s1997 xx |||||o 00| ||eng c | ||
| 024 | 7 | |a 10.2144/97223st05 |2 doi | |
| 035 | |a (DE-627)1914059867 | ||
| 035 | |a (DE-599)KXP1914059867 | ||
| 035 | |a (OCoLC)1528015921 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rda | ||
| 041 | |a eng | ||
| 084 | |a 32 |2 sdnb | ||
| 100 | 1 | |a Zhan, Jinghai |d 1961- |e VerfasserIn |0 (DE-588)115645675 |0 (DE-627)691478465 |0 (DE-576)289997291 |4 aut | |
| 245 | 1 | 0 | |a Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA |c Jinghai Zhan, H. Dariush Fahimi, Alfred Völkl |
| 264 | 1 | |c 1997 | |
| 300 | |a 6 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 500 | |a Elektronische Reproduktion der Druck-Ausgabe 7. August 2018 | ||
| 500 | |a Gesehen am 09.01.2025 | ||
| 520 | |a We have developed a simple and sensitive method for the rapid quantitation of mRNA from cell cultures and small tissue samples. The method combines the high sensitivity and specificity of the ribonuclease protection assay with simple handling and rapid execution of dot blotting. The use of digoxygenin-labeled cRNA probes eliminates all problems associated with radioisotopes commonly used in the ribonuclease protection assay. The RNA preparation is dotted directly onto nylon membranes, and after hybridization the filters are treated with ribonuclease A, which removes the nonhybridized single-stranded RNA. The mRNA-hybrid is then visualized by the chemiluminescence technique using labeled anti-digoxigenin antibody, and the signal intensity is quantitated. Comparison with the Northern blotting ribonuclease protection assay revealed that this dot blot technique is almost ten times more sensitive and that its signals are linear over a wide range of RNA concentrations (0.01-10 μg/μL/ dot). This method seems particularly valuable for simultaneous processing of large numbers of samples containing a wide range of RNA concentrations. | ||
| 700 | 1 | |a Fahimi, H. Dariush |d 1933- |e VerfasserIn |0 (DE-588)1114523305 |0 (DE-627)868737194 |0 (DE-576)164247742 |4 aut | |
| 700 | 1 | |a Völkl, Alfred |d 1940- |e VerfasserIn |0 (DE-588)1021086045 |0 (DE-627)691446970 |0 (DE-576)362512140 |4 aut | |
| 773 | 0 | 8 | |i Enthalten in |t BioTechniques |d London, UK : Future Science Ltd, 1983 |g 22(1997), 3, Seite 500-505 |h Online-Ressource |w (DE-627)306320746 |w (DE-600)1496354-1 |w (DE-576)094115559 |x 1940-9818 |7 nnas |a Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA |
| 773 | 1 | 8 | |g volume:22 |g year:1997 |g number:3 |g pages:500-505 |g extent:6 |a Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA |
| 856 | 4 | 0 | |u https://doi.org/10.2144/97223st05 |x Verlag |x Resolving-System |z lizenzpflichtig |3 Volltext |
| 951 | |a AR | ||
| 992 | |a 20250109 | ||
| 993 | |a Article | ||
| 994 | |a 1997 | ||
| 998 | |g 1021086045 |a Völkl, Alfred |m 1021086045:Völkl, Alfred |d 50000 |d 53100 |e 50000PV1021086045 |e 53100PV1021086045 |k 0/50000/ |k 1/50000/53100/ |p 3 |y j | ||
| 998 | |g 1114523305 |a Fahimi, H. Dariush |m 1114523305:Fahimi, H. Dariush |d 50000 |d 53100 |e 50000PF1114523305 |e 53100PF1114523305 |k 0/50000/ |k 1/50000/53100/ |p 2 | ||
| 998 | |g 115645675 |a Zhan, Jinghai |m 115645675:Zhan, Jinghai |d 50000 |d 53100 |e 50000PZ115645675 |e 53100PZ115645675 |k 0/50000/ |k 1/50000/53100/ |p 1 |x j | ||
| 999 | |a KXP-PPN1914059867 |e 4648241665 | ||
| BIB | |a Y | ||
| SER | |a journal | ||
| JSO | |a {"id":{"eki":["1914059867"],"doi":["10.2144/97223st05"]},"origin":[{"dateIssuedDisp":"1997","dateIssuedKey":"1997"}],"name":{"displayForm":["Jinghai Zhan, H. Dariush Fahimi, Alfred Völkl"]},"relHost":[{"title":[{"title_sort":"BioTechniques","subtitle":"the international journal of life science methods","title":"BioTechniques"}],"language":["eng"],"recId":"306320746","disp":"Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNABioTechniques","type":{"media":"Online-Ressource","bibl":"periodical"},"note":["Gesehen am 30.05.2024","Frühere Jahrgänge online nicht mehr verfügbar"],"part":{"extent":"6","text":"22(1997), 3, Seite 500-505","volume":"22","pages":"500-505","issue":"3","year":"1997"},"pubHistory":["1.1983 -"],"id":{"issn":["1940-9818"],"zdb":["1496354-1"],"eki":["306320746"]},"origin":[{"publisher":"Future Science Ltd ; Eaton Publ. Co. ; Eaton ; Informa Life Sciences Publ.","dateIssuedKey":"1983","dateIssuedDisp":"1983-","publisherPlace":"London, UK ; Natick, Mass. ; Westborough, Mass. ; Westborough, Mass."}],"physDesc":[{"extent":"Online-Ressource"}]}],"physDesc":[{"extent":"6 S."}],"title":[{"title_sort":"Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA","title":"Sensitive nonradioactive dot blot/ribonuclease protection assay for quantitative determination of mRNA"}],"person":[{"given":"Jinghai","family":"Zhan","role":"aut","display":"Zhan, Jinghai","roleDisplay":"VerfasserIn"},{"role":"aut","roleDisplay":"VerfasserIn","display":"Fahimi, H. Dariush","given":"H. Dariush","family":"Fahimi"},{"given":"Alfred","family":"Völkl","role":"aut","display":"Völkl, Alfred","roleDisplay":"VerfasserIn"}],"language":["eng"],"recId":"1914059867","type":{"media":"Online-Ressource","bibl":"article-journal"},"note":["Elektronische Reproduktion der Druck-Ausgabe 7. August 2018","Gesehen am 09.01.2025"]} | ||
| SRT | |a ZHANJINGHASENSITIVEN1997 | ||