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Quantification of microRNAs and DNA methylation status by qPCR

Published on Jun 01, 20151853 Views

MicroRNAs and DNA methylation are both critical epigenetic mechanisms for the control of gene expression. The recent explosion of interest in these two areas has led to the development of innovative s

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Molecular Biology

Chapter list

Quantification of microRNAs and DNA methylation status by qPCR00:00
Overview00:13
miRNA biogenesis01:13
miRNAs02:55
Functions of miRNAs04:35
miRNAs: Small molecules with a big impact05:33
miRNA quantification platforms07:37
Challenges for miRNA qPCR08:19
miRNA RT-qPCR technologies09:35
Stem-loop RT-qPCR: Life Technologies10:30
Universal RT priming: Exiqon12:04
miScript PCR System (Qiagen)14:12
Summary of miRNA qPCR methods15:46
miRNA qPCR specificity16:20
miRNA qPCR normalisation17:27
miRNA RT-qPCR arrays - 118:48
miRNA RT-qPCR arrays - 219:25
miRNA RT-qPCR arrays - 319:52
Obtaining accurate and consistent miRNA data20:57
miRNA extraction methods - 121:21
miRNA extraction methods - 222:31
Extraction yield23:24
Total RNA vs. short RNA enrichment?24:37
miRNA qPCR assay performance26:00
Comparison of extraction and qPCR technologies for miRNA quantification27:10
Measurement of miRNA spike-ins31:43
Part 2: Quantification of DNA methylation status by qPCR32:07
Epigenetics32:09
DNA methylation32:56
Gene Regulation: DNA methylation in diagnostics35:55
Methods and platforms37:04
Methylation quantification by PCR38:05
Bisulfite conversion of unmethylated Cytosine residues39:06
Bisulfite conversion40:15
Bisulfite PCR40:39
Methylight qPCR42:10
Bisulfite PCR primer design42:58
Bisulfite PCR – warning!44:55
Methylight qPCR performance46:09
LGC Methylight qPCR analysis48:17
LGC Methylight qPCR analysis48:50
Methylight qPCR: Normalisation50:00
Methylight qPCR: Bias51:19
Methylight qPCR54:19
Quantification of methylation using Restriction Enzyme qPCR - 155:26
Quantification of methylation using Restriction Enzyme qPCR - 258:56
Quantification of methylation using Restriction Enzyme qPCR - 359:32
LGC Restriction Enzyme qPCR analysis59:52
Restriction enzyme qPCR01:00:42
Combined Bisulfite Restriction Analysis (COBRA)01:02:37
COBRA analysis01:03:33
COBRA01:04:19
Summary01:05:06
Thank you01:05:46