The Experts below are selected from a list of 161559 Experts worldwide ranked by ideXlab platform
M C Wiltbank - One of the best experts on this subject based on the ideXlab platform.
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quantification of mrna using competitive rt pcr with Standard Curve methodology
BioTechniques, 1996Co-Authors: Shaw Jenq Tsai, M C WiltbankAbstract:The use of reverse transcription polymerase chain reaction (RT-PCR) with internal RNA competitive Standards (competitors) provides a means for measuring absolute amounts of mRNA transcripts in smal...
Robyn M. Murphy - One of the best experts on this subject based on the ideXlab platform.
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are genuine changes in protein expression being overlooked reassessing western blotting
Analytical Biochemistry, 2009Co-Authors: Janelle P Mollica, G D Lamb, Johathan S Oakhill, Robyn M. MurphyAbstract:This study used purified calsequestrin 1 and AMP kinase (AMPK) proteins to demonstrate how Western blotting outcomes can be influenced when either the density of proteins detected lie within a nonproportional region of a Standard Curve or a Standard Curve is not taken into account for data analyses. It outlines the likelihood of true changes being overlooked through the simple mistake of using band density alone and/or through analyzing too much sample. To demonstrate this, extrapolation of a typical linear, although nonproportional, Standard Curve resulted in approximately fourfold error. The Standard Curve method was used to estimate the concentration of AMPK β1 in rat extensor digitorum longus muscle as being of the order of 60 μM. The article suggests that adopting a more sensitive Western blotting protocol will improve the reliability of quantitative Western blotting outcomes.
Neville A Mcbrien - One of the best experts on this subject based on the ideXlab platform.
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high resolution semi quantitative real time pcr without the use of a Standard Curve
BioTechniques, 2001Co-Authors: Alex Gentle, Frank Anastasopoulos, Neville A McbrienAbstract:The repeatability and sensitivity of a simple, adaptable, semi-quantitative, real-time RT-PCR assay was investigated. The assay can be easily and rapidly applied to quantitate relative levels of any gene product without using Standards, provided that amplification conditions are specific for the PCR product of interest. Using the LightCycler real-time PCR machine, a serial 10-fold dilution series (spanning four orders of magnitude) of a 379-bp cDNA template was amplified, and the PCR product was detected using SYBR Green I chemistry. The experiment was repeated on a subsequent day. The experimental design was such that the data lent itself to analysis using an appropriate method for testing repeatability. It was found that, within a single assay, for samples assayed in triplicate, a difference of 23% may be reliably detected. Furthermore, when all of the factors that contribute to variability in the assay are taken into account, such as day-to-day variation in pipetting and amplification efficiency, a 52% difference in target template can be detected using a sample size of 4. The assay was found to be linear over at least four orders of magnitude.
Shaw Jenq Tsai - One of the best experts on this subject based on the ideXlab platform.
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quantification of mrna using competitive rt pcr with Standard Curve methodology
BioTechniques, 1996Co-Authors: Shaw Jenq Tsai, M C WiltbankAbstract:The use of reverse transcription polymerase chain reaction (RT-PCR) with internal RNA competitive Standards (competitors) provides a means for measuring absolute amounts of mRNA transcripts in smal...
Janelle P Mollica - One of the best experts on this subject based on the ideXlab platform.
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are genuine changes in protein expression being overlooked reassessing western blotting
Analytical Biochemistry, 2009Co-Authors: Janelle P Mollica, G D Lamb, Johathan S Oakhill, Robyn M. MurphyAbstract:This study used purified calsequestrin 1 and AMP kinase (AMPK) proteins to demonstrate how Western blotting outcomes can be influenced when either the density of proteins detected lie within a nonproportional region of a Standard Curve or a Standard Curve is not taken into account for data analyses. It outlines the likelihood of true changes being overlooked through the simple mistake of using band density alone and/or through analyzing too much sample. To demonstrate this, extrapolation of a typical linear, although nonproportional, Standard Curve resulted in approximately fourfold error. The Standard Curve method was used to estimate the concentration of AMPK β1 in rat extensor digitorum longus muscle as being of the order of 60 μM. The article suggests that adopting a more sensitive Western blotting protocol will improve the reliability of quantitative Western blotting outcomes.
