Get tips on using QuantiPro™ BCA Assay Kit to perform Protein quantification Mammalian cells - RAW264.7
Get tips on using Qubit™ Protein Assay Kit to perform Protein quantification Mammalian cells - HeLa
Get tips on using SQSTM1/p62 Antibody #5114 to perform Immunohistochemistry Mouse - p62
Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Rat skin tissue
Get tips on using CelLytic™ MT Cell Lysis Reagent to perform Protein isolation Tissue - Mouse skeletal muscle
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 CellTiter-Glo® Luminescent Cell Viability Assay to perform Live / Dead assay mammalian cells - H9C2
Get tips on using CellTiter-Glo® Luminescent Cell Viability Assay to perform Live / Dead assay mammalian cells - MMQ
Get tips on using CellTiter-Glo® Luminescent Cell Viability Assay to perform Live / Dead assay mammalian cells - GH3
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