Get tips on using MethylCap kit to perform DNA methylation profiling Whole genome profiling - HCT116, HTC15 human colon cancer cells
Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Whole genome profiling - C2C12 mouse myoblast cells
Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Whole genome profiling - mouse primordial germ cells
Get tips on using Hydroxymethyl Collector™ Kit to perform DNA methylation profiling Whole genome profiling - mouse primordial germ cells
Get tips on using EZ DNA Methylation kit to perform DNA methylation profiling Whole genome profiling - mouse hematopoietic stem cells
Get tips on using SurePrint G3 Human CGH Microarray Kit, 4x180K to perform Microarray Comperative genomic hybridization - Human Blood cells
Get tips on using SurePrint G3 Human CGH Microarray Kit, 2x400K to perform Microarray Comperative genomic hybridization - Human Blood cells
The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.
The formation of DNA from an RNA template using reverse transcription leads to the formation of double-stranded complementary DNA or cDNA. The challenges with this process include 1. Maintaining the integrity of RNA, 2. Hairpin loops or other secondary structures formed by single-stranded RNA can also affect cDNA synthesis, and 3. DNA-RNA hybrids, which may result when the first strand of cDNA is formed. For the first challenge, using workflows that involve proper isolation and storage of RNA, and maintaining a nuclease-free environment helps obtain RNA with ideal 260/230 ratios. Using a reverse transcriptase that can tolerate high temperatures (50-55oC), overcomes obstacles imposed by secondary RNA structures. Finally, RNase H has the ability to hydrolyze RNA before the formation of a second cDNA strand. It is important to ensure that RNase H activity is optimal because higher RNase H activity leads to premature degradation of the RNA template. Many reverse transcriptases offer built-in RNase H activity.
Get tips on using CytoTox 96® Non-Radioactive Cytotoxicity Assay to perform Live / Dead assay mammalian cells - mouse bone marrow-derived macrophages
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