Semiquantitative RT-PCR: Enhancement of Assay Accuracy and Reproducibility
نویسندگان
چکیده
منابع مشابه
Reproducibility in the quantification of mRNA levels by RT-PCR-ELISA and RT competitive-PCR-ELISA.
The use of reverse transcription (RT) PCR for relative quantitation of gene transcripts relies on the reproducibility of the individual RT, PCR and product measurement steps. Semi-competitive RT-PCR (RT-cPCR) uses an internal competitor template in the PCR step to improve quantitation. We have surveyed the reproducibility of RT, PCR, RT-cPCR and measurement, amplifying the glyceraldehyde-3-phos...
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In this paper we determine the limits and accuracy of quantitative reverse transcription (RT)-PCR using a modification of the original protocol. The quantification of mRNA with this procedure requires a preliminary estimation of the target molecule (TM) concentration, established from experiments with an internal control molecule (ICM). A definitive quantification is then attained from serial d...
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Abstract: iNOS is inducible by a variety of factors related to inflammation and referred to as inducible NOS(iNOS). It is regulated at the level of gene expression; once expressed, it produces NO at a high rate. iNOS gene-expression profiling is an important tool in understanding molecular markers of the responses of cells and tissues to external factors. In this article a semiquantitative reve...
متن کاملInvestigation of beta defensin gene expression in the ocular anterior segment by semiquantitative RT-PCR.
AIM To determine if beta defensins are expressed in the anterior segment of the eye and to determine the temporal pattern of expression using a real time semiquantitative reverse transcription polymerase chain reaction (RT-PCR). METHODS Ocular tissue (corneal epithelium, conjunctiva, iris, and lens capsule) was collected from 23 patients undergoing surgery. Serial corneal or conjunctival impr...
متن کاملInvestigation of â defensin gene expression in the ocular anterior segment by semiquantitative RT-PCR
Aim—To determine if â defensins are expressed in the anterior segment of the eye and to determine the temporal pattern of expression using a real time semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Methods—Ocular tissue (corneal epithelium, conjunctiva, iris, and lens capsule) was collected from 23 patients undergoing surgery. Serial corneal or conjunctival impressio...
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ژورنال
عنوان ژورنال: BioTechniques
سال: 1997
ISSN: 0736-6205,1940-9818
DOI: 10.2144/97224bm12