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High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis

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dc.contributor.author Van den Berg, N
dc.contributor.author Crampton, BG
dc.contributor.author Hein, I
dc.contributor.author Birch, PRJ
dc.contributor.author Berger, DK
dc.date.accessioned 2007-06-12T07:20:10Z
dc.date.available 2007-06-12T07:20:10Z
dc.date.issued 2004-11
dc.identifier.citation Van den Berg, N, et al. 2004. High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis. Biotechniques, vol. 37 (5), pp 818-824 en
dc.identifier.issn 0736-6205
dc.identifier.uri http://hdl.handle.net/10204/523
dc.description Copyright: 2004 Eaton Publishing Co en
dc.description.abstract Efficient construction of cDNA libraries enriched for differentially expressed transcripts is an important first step in many biological investigations. We present a quantitative procedure for screening cDNA libraries constructed by suppression subtractive hybridization (SSH). The methodology was applied to two independent SSHs from pearl millet and banana. Following two-colour cyanin dye labelling and hybridization of subtracted tester with either unsubtracted driver or unsubtracted tester cDNAs to the SSH libraries arrayed on glass slides, two values were calculated for each clone, an enrichment ratio 1 (ER1) and an enrichment ratio 2 (ER2). Graphical representation of ER1 and ER2 enabled the identification of clones that were likely to represent up-regulated transcripts. Normalization of each clone by the SSH process was determined from the ER2 values, thereby indicating whether clones represented rare or abundant transcripts. Differential expression of pearl millet and banana clones identified from both libraries by this quantitative approach was verified by inverse Northern blot analysis. en
dc.language.iso en en
dc.publisher Eaton Publishing Co en
dc.subject cDNA synthesis en
dc.subject DNA macroarray analysis en
dc.subject Subtractive hybridizations en
dc.subject Plant treatments en
dc.title High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis en
dc.type Article en
dc.identifier.apacitation Van den Berg, N., Crampton, B., Hein, I., Birch, P., & Berger, D. (2004). High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis. http://hdl.handle.net/10204/523 en_ZA
dc.identifier.chicagocitation Van den Berg, N, BG Crampton, I Hein, PRJ Birch, and DK Berger "High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis." (2004) http://hdl.handle.net/10204/523 en_ZA
dc.identifier.vancouvercitation Van den Berg N, Crampton B, Hein I, Birch P, Berger D. High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis. 2004; http://hdl.handle.net/10204/523. en_ZA
dc.identifier.ris TY - Article AU - Van den Berg, N AU - Crampton, BG AU - Hein, I AU - Birch, PRJ AU - Berger, DK AB - Efficient construction of cDNA libraries enriched for differentially expressed transcripts is an important first step in many biological investigations. We present a quantitative procedure for screening cDNA libraries constructed by suppression subtractive hybridization (SSH). The methodology was applied to two independent SSHs from pearl millet and banana. Following two-colour cyanin dye labelling and hybridization of subtracted tester with either unsubtracted driver or unsubtracted tester cDNAs to the SSH libraries arrayed on glass slides, two values were calculated for each clone, an enrichment ratio 1 (ER1) and an enrichment ratio 2 (ER2). Graphical representation of ER1 and ER2 enabled the identification of clones that were likely to represent up-regulated transcripts. Normalization of each clone by the SSH process was determined from the ER2 values, thereby indicating whether clones represented rare or abundant transcripts. Differential expression of pearl millet and banana clones identified from both libraries by this quantitative approach was verified by inverse Northern blot analysis. DA - 2004-11 DB - ResearchSpace DP - CSIR KW - cDNA synthesis KW - DNA macroarray analysis KW - Subtractive hybridizations KW - Plant treatments LK - https://researchspace.csir.co.za PY - 2004 SM - 0736-6205 T1 - High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis TI - High-throughput screening of suppression subtractive hybridization cDNA libraries using DNA microarray analysis UR - http://hdl.handle.net/10204/523 ER - en_ZA


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