Differential Display Probes for cDNA Arrays
PCR with a combination of one arbitrary and one oligo(dT) anchor primer can be used to generate an effective probe for cDNA arrays. The method uses less than 1/200 of the amount of RNA used in some other array hybridization methods. Each fingerprint detects approximately 5% of the transcribed mRNAs,...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
1999-09-01
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| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/99273rr03 |
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| author | Thomas Trenkle John Welsh Michael McClelland |
| author_facet | Thomas Trenkle John Welsh Michael McClelland |
| author_sort | Thomas Trenkle |
| collection | DOAJ |
| description | PCR with a combination of one arbitrary and one oligo(dT) anchor primer can be used to generate an effective probe for cDNA arrays. The method uses less than 1/200 of the amount of RNA used in some other array hybridization methods. Each fingerprint detects approximately 5% of the transcribed mRNAs, sampled almost independent of abundance, using inexpensive E. coli colony arrays of expressed sequence tag (EST) clones. It proved necessary to alter the differential display (DD) protocol to generate a sufficient mass of PCR products for use as a probe. The use of different oligo( dT) anchor primers with the same arbitrary primer resulted in considerable overlap among the genes sampled by each probe. This can be avoided by using different arbitrary primers with each oligo(dT) anchor primer. Four genes not previously known to be regulated by epidermal growth factor (EGF) and three genes known to be regulated by EGF in other cell types were characterized using DD fingerprints as probes for arrays. It should be possible to convert archived DD fingerprints into effective probes for arrays, allowing thousands of experiments that have already been performed to yield further information. The use of DD fingerprints as probes should increase the rate of identification of differentially regulated genes several fold while obviating the need for cloning and sequencing. |
| format | Article |
| id | doaj-art-602838f64d0645a4bcc26fdee114438e |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 1999-09-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-602838f64d0645a4bcc26fdee114438e2025-08-20T02:25:34ZengTaylor & Francis GroupBioTechniques0736-62051940-98181999-09-0127355456410.2144/99273rr03Differential Display Probes for cDNA ArraysThomas Trenkle0John Welsh1Michael McClelland21Sidney Kimmel Cancer Center, San Diego, CA, USA1Sidney Kimmel Cancer Center, San Diego, CA, USA1Sidney Kimmel Cancer Center, San Diego, CA, USAPCR with a combination of one arbitrary and one oligo(dT) anchor primer can be used to generate an effective probe for cDNA arrays. The method uses less than 1/200 of the amount of RNA used in some other array hybridization methods. Each fingerprint detects approximately 5% of the transcribed mRNAs, sampled almost independent of abundance, using inexpensive E. coli colony arrays of expressed sequence tag (EST) clones. It proved necessary to alter the differential display (DD) protocol to generate a sufficient mass of PCR products for use as a probe. The use of different oligo( dT) anchor primers with the same arbitrary primer resulted in considerable overlap among the genes sampled by each probe. This can be avoided by using different arbitrary primers with each oligo(dT) anchor primer. Four genes not previously known to be regulated by epidermal growth factor (EGF) and three genes known to be regulated by EGF in other cell types were characterized using DD fingerprints as probes for arrays. It should be possible to convert archived DD fingerprints into effective probes for arrays, allowing thousands of experiments that have already been performed to yield further information. The use of DD fingerprints as probes should increase the rate of identification of differentially regulated genes several fold while obviating the need for cloning and sequencing.https://www.future-science.com/doi/10.2144/99273rr03 |
| spellingShingle | Thomas Trenkle John Welsh Michael McClelland Differential Display Probes for cDNA Arrays BioTechniques |
| title | Differential Display Probes for cDNA Arrays |
| title_full | Differential Display Probes for cDNA Arrays |
| title_fullStr | Differential Display Probes for cDNA Arrays |
| title_full_unstemmed | Differential Display Probes for cDNA Arrays |
| title_short | Differential Display Probes for cDNA Arrays |
| title_sort | differential display probes for cdna arrays |
| url | https://www.future-science.com/doi/10.2144/99273rr03 |
| work_keys_str_mv | AT thomastrenkle differentialdisplayprobesforcdnaarrays AT johnwelsh differentialdisplayprobesforcdnaarrays AT michaelmcclelland differentialdisplayprobesforcdnaarrays |