| pubmed-article:12753500 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C0025663 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C0032520 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C0017262 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C1333196 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C1705938 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C1527178 | lld:lifeskim |
| pubmed-article:12753500 | lifeskim:mentions | umls-concept:C0386577 | lld:lifeskim |
| pubmed-article:12753500 | pubmed:issue | 5 | lld:pubmed |
| pubmed-article:12753500 | pubmed:dateCreated | 2003-5-19 | lld:pubmed |
| pubmed-article:12753500 | pubmed:abstractText | Assessment of cytokine expression has become crucial to understand host responses to infections as well as autoimmunity. Several approaches including Northern blot, RNase protection assay and enzyme-linked immunosorbent assay have been used for this purpose, but they are time consuming, labour intense, and relatively large quantity of the samples is usually required. Recently, a technique termed real-time reverse transcriptase-polymerase chain reaction (RT-PCR) has been developed to determine genetic expression with great sensitivity and specificity; however, specialized instrumentation and costly reagents are usually needed. We aimed at using low-cost reagents for real-time PCR. This was achieved by adapting a conventional RT-PCR protocol to the quantitative real-time format, by the addition of the SYBR Green I reagent. We validated the approach by assessing the cytokine gene expression of murine splenocytes upon stimulation with phorbol 12-myristate 12-acetate (PMA)-ionomycin. The results using this technique were compared with those obtained with the well-established gene array method. We conclude that the use of the SYBR Green I reagent during real-time RT-PCR provides a highly specific and sensitive method to quantify cytokine expression with accuracy and no post-PCR manipulation. | lld:pubmed |
| pubmed-article:12753500 | pubmed:language | eng | lld:pubmed |
| pubmed-article:12753500 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:12753500 | pubmed:citationSubset | IM | lld:pubmed |
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| pubmed-article:12753500 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:12753500 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:12753500 | pubmed:month | May | lld:pubmed |
| pubmed-article:12753500 | pubmed:issn | 0300-9475 | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Estrada-Parra... | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Favila-Castil... | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Monroy-Ostria... | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Aguilar-Medin... | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Ramos-PayánRR | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:González-Y-Me... | lld:pubmed |
| pubmed-article:12753500 | pubmed:author | pubmed-author:Estrada-Garci... | lld:pubmed |
| pubmed-article:12753500 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:12753500 | pubmed:volume | 57 | lld:pubmed |
| pubmed-article:12753500 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:12753500 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:12753500 | pubmed:pagination | 439-45 | lld:pubmed |
| pubmed-article:12753500 | pubmed:dateRevised | 2008-11-21 | lld:pubmed |
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| pubmed-article:12753500 | pubmed:year | 2003 | lld:pubmed |
| pubmed-article:12753500 | pubmed:articleTitle | Quantification of cytokine gene expression using an economical real-time polymerase chain reaction method based on SYBR Green I. | lld:pubmed |
| pubmed-article:12753500 | pubmed:affiliation | Departamento de Inmunología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional, México D F, México. | lld:pubmed |
| pubmed-article:12753500 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:12753500 | pubmed:publicationType | Research Support, Non-U.S. Gov't | lld:pubmed |
| pubmed-article:12753500 | pubmed:publicationType | Evaluation Studies | lld:pubmed |
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