Search results for "cDNA synthesis"

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Normalization with Corresponding Naïve Tissue Minimizes Bias Caused by Commercial Reverse Transcription Kits on Quantitative Real-Time PCR Results

2016

Real-time reverse transcription polymerase chain reaction (PCR) is the gold standard for expression analysis. Designed to improve reproducibility and sensitivity, commercial kits are commonly used for the critical step of cDNA synthesis. The present study was designed to determine the impact of these kits. mRNA from mouse brains were pooled to create serial dilutions ranging from 0.0625 μg to 2 μg, which were transcribed into cDNA using four different commercial reverse-transcription kits. Next, we transcribed mRNA from brain tissue after acute brain injury and naïve mice into cDNA for qPCR. Depending on tested genes, some kits failed to show linear results in dilution series and revealed s…

Male0301 basic medicineSerial dilutionlcsh:MedicineGene ExpressioncDNA synthesisArtificial Gene Amplification and ExtensionBioinformaticsBiochemistryPolymerase Chain ReactionMice0302 clinical medicineBrain Injuries Traumaticlcsh:ScienceGenes EssentialMultidisciplinaryReverse Transcriptase Polymerase Chain ReactionMessenger RNAComplementary DNAHousekeeping geneNucleic acidsReverse transcription polymerase chain reactionResearch ArticleNormalization (statistics)DNA ComplementaryForms of DNANucleic acid synthesisBiologyReal-Time Polymerase Chain ReactionResearch and Analysis Methods03 medical and health sciencesExtraction techniquesComplementary DNAGeneticsAnimalsRNA MessengerChemical synthesisRNA synthesisMolecular Biology TechniquesMolecular BiologyGeneMessenger RNABiology and life scienceslcsh:RDNAReverse TranscriptionMolecular biologyRNA extractionReverse transcriptaseMice Inbred C57BLBiosynthetic techniques030104 developmental biologyRNAlcsh:Q030217 neurology & neurosurgeryPLOS ONE

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PEPhysical Sciences and Engineering
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LSLife Sciences
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