Abstract
Expressed Sequence Tags (ESTs) provide a rapid and efficient approach for gene discovery and analysis of gene expression in eukaryotes. ESTs have also become particularly important with recent expanded efforts in complete genome sequencing of understudied, nonmodel eukaryotes such as protists and algae. For these projects, ESTs provide an invaluable source of data for gene identification and prediction of exon-intron boundaries. The generation of EST data, although straightforward in concept, requires nonetheless great care to ensure the highest efficiency and return for the investment in time and funds. To this end, key steps in the process include generation of a normalized cDNA library to facilitate a high gene discovery rate followed by serial subtraction of normalized libraries to maintain the discovery rate. Here we describe in detail, protocols for normalization and subtraction of cDNA libraries followed by an example using the toxic dinoflagellate Alexandrium tamarense.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 109-122 |
| Number of pages | 14 |
| Journal | Methods in molecular biology (Clifton, N.J.) |
| Volume | 533 |
| State | Published - 2009 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Molecular Biology
- Genetics
Cite this
Expressed sequence tags : normalization and subtraction of cDNA libraries expressed sequence tags\ normalization and subtraction of cDNA libraries. / Soares, Marcelo Bento; de Fatima Bonaldo, Maria; Hackett, Jeremiah D.; Bhattacharya, Debashish.
In: Methods in molecular biology (Clifton, N.J.), Vol. 533, 2009, p. 109-122.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Expressed sequence tags
T2 - normalization and subtraction of cDNA libraries expressed sequence tags\ normalization and subtraction of cDNA libraries.
AU - Soares, Marcelo Bento
AU - de Fatima Bonaldo, Maria
AU - Hackett, Jeremiah D.
AU - Bhattacharya, Debashish
PY - 2009
Y1 - 2009
N2 - Expressed Sequence Tags (ESTs) provide a rapid and efficient approach for gene discovery and analysis of gene expression in eukaryotes. ESTs have also become particularly important with recent expanded efforts in complete genome sequencing of understudied, nonmodel eukaryotes such as protists and algae. For these projects, ESTs provide an invaluable source of data for gene identification and prediction of exon-intron boundaries. The generation of EST data, although straightforward in concept, requires nonetheless great care to ensure the highest efficiency and return for the investment in time and funds. To this end, key steps in the process include generation of a normalized cDNA library to facilitate a high gene discovery rate followed by serial subtraction of normalized libraries to maintain the discovery rate. Here we describe in detail, protocols for normalization and subtraction of cDNA libraries followed by an example using the toxic dinoflagellate Alexandrium tamarense.
AB - Expressed Sequence Tags (ESTs) provide a rapid and efficient approach for gene discovery and analysis of gene expression in eukaryotes. ESTs have also become particularly important with recent expanded efforts in complete genome sequencing of understudied, nonmodel eukaryotes such as protists and algae. For these projects, ESTs provide an invaluable source of data for gene identification and prediction of exon-intron boundaries. The generation of EST data, although straightforward in concept, requires nonetheless great care to ensure the highest efficiency and return for the investment in time and funds. To this end, key steps in the process include generation of a normalized cDNA library to facilitate a high gene discovery rate followed by serial subtraction of normalized libraries to maintain the discovery rate. Here we describe in detail, protocols for normalization and subtraction of cDNA libraries followed by an example using the toxic dinoflagellate Alexandrium tamarense.
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M3 - Article
VL - 533
SP - 109
EP - 122
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
SN - 1064-3745
ER -