rna-isolation-purification-tissue-rat-stomach

Get tips on using EasySep™ Human Naïve B Cell Isolation Kit to perform Cell Isolation Naive B cell

Get tips on using EasySep™ Direct Human T Cell Isolation Kit to perform Cell Isolation Human T cells

Get tips on using MACSprep™ HLA B Cell Isolation Kit, human to perform Cell Isolation HLA B Cell

Get tips on using MACSprep™ HLA T Cell Isolation Kit, human to perform Cell Isolation HLA T Cell

Microarrays enable researchers to monitor the expression of thousands of genes simultaneously. However, the sensitivity, accuracy, specificity, and reproducibility are major challenges for this technology. Cross-hybridization, combination with splice variants, is a prime source for the discrepancies in differential gene expression calls among various microarray platforms. Removing (either from production or downstream bioinformatic analysis) and/or redesigning the microarray probes prone to cross-hybridization is a reasonable strategy to increase the hybridization specificity and hence, the accuracy of the microarray measurements.

Get tips on using TIANamp Genomic DNA Kit to perform DNA isolation / purification Cells - Primary cells Mouse embryonic fibroblast (MEF)

Get tips on using Gentra Puregene Blood Kit to perform DNA isolation / purification Cells - Primary cells Bone marrow mononuclear cells

Get tips on using Gentra Puregene Blood Kit to perform DNA isolation / purification Cells - Immortalized cell lines Lymphoblastoid cell lines

Get tips on using QIAamp DNA Blood Mini Kit to perform DNA isolation / purification Cells - Immortalized cell lines HEK 293T

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.