Gene Expression Analysis by Reverse Transcription Quantitative PCR

2021 ◽  
pp. 61-74
Author(s):  
Eva M. Campion ◽  
Sinéad T. Loughran
Keyword(s):  
Gene Expression ◽  
Quantitative Pcr ◽  
Expression Analysis ◽  
Reverse Transcription ◽  
Gene Expression Analysis ◽  
Reverse Transcription Quantitative Pcr

scholarly journals Reliable Gene Expression Analysis by Reverse Transcription-Quantitative PCR: Reporting and Minimizing the Uncertainty in Data Accuracy

2014 ◽  
Vol 26 (10) ◽  
pp. 3829-3837 ◽  
Author(s):  
Tony Remans ◽  
Els Keunen ◽  
Geert Jan Bex ◽  
Karen Smeets ◽  
Jaco Vangronsveld ◽  
...  
Keyword(s):  
Gene Expression ◽  
Quantitative Pcr ◽  
Expression Analysis ◽  
Reverse Transcription ◽  
Gene Expression Analysis ◽  
Data Accuracy ◽  
Reverse Transcription Quantitative Pcr

scholarly journals Maximizing RNA Yield from Archival Renal Tumors and Optimizing Gene Expression Analysis

2009 ◽  
Vol 15 (1) ◽  
pp. 80-85 ◽  
Author(s):  
Sean T. Glenn ◽  
Karen L. Head ◽  
Bin T. Teh ◽  
Kenneth W. Gross ◽  
Hyung L. Kim
Keyword(s):  
Gene Expression ◽  
Expression Analysis ◽  
Reverse Transcription ◽  
Gene Expression Analysis ◽  
Rna Isolation ◽  
Proteinase K ◽  
Specific Primers ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Formalin-fixed, paraffin-embedded tissues are widely available for gene expression analysis using TaqMan ® PCR. Five methods, including 4 commercial kits, for recovering RNA from paraffin-embedded renal tumor tissue were compared. The MasterPure™ kit from Epicentre produced the highest RNA yield. However, the difference in RNA yield between the kit from Epicenter and Invitrogen’s TRIzol method was not significant. Using the top 3 RNA isolation methods, the manufacturers’ protocols were modified to include an overnight Proteinase K digestion. Overnight protein digestion resulted in a significant increase in RNA yield. To optimize the reverse transcription reaction, conventional reverse transcription with random oligonucleotide primers was compared to reverse transcription using primers specific for genes of interest. Reverse transcription using gene-specific primers significantly increased the quantity of cDNA detectable by TaqMan ® PCR. Therefore, expression profiling of formalin-fixed, paraffin-embedded tissue using TaqMan® qPCR can be optimized by using the MasterPure™ RNA isolation kit modified to include an overnight Proteinase K digestion and gene-specific primers during the reverse transcription.

scholarly journals Twenty-five years of quantitative PCR for gene expression analysis

BioTechniques ◽  
2008 ◽  
Vol 44 (5) ◽  
pp. 619-626 ◽  
Author(s):  
Heather D. VanGuilder ◽  
Kent E. Vrana ◽  
Willard M. Freeman
Keyword(s):  
Gene Expression ◽  
Quantitative Pcr ◽  
Expression Analysis ◽  
Gene Expression Analysis
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