siRNA / miRNA gene silencing Human 501 Mel and SK Mel 28 FANCD2

I am currently using a recombinant protein which shows metal-dependent DNase activity. Is it possible to pinpoint the source of the DNase activity after protein purification? More specifically, can I ensure that the DNase activity is not because of nuclease contamination from the E.coli that might have persisted and passed with the protein of interest during purification?

Get tips on using GeneJET Genomic DNA Purification Kit to perform DNA isolation / purification Bacteria - Gram positive Streptomyces. Sp

Get tips on using Brilliant Violet 510™ anti-mouse CD69 Antibody to perform Flow cytometry Anti-bodies Mouse - CD69

Get tips on using GenElute™ Bacterial Genomic DNA Kit to perform DNA isolation / purification Bacteria - Gram positive Actinomycytes

Get tips on using GenElute™ Bacterial Genomic DNA Kit to perform DNA isolation / purification Bacteria - Gram negative E.coli

Get tips on using Luria Bertani Broth, Miller (Miller Luria Bertani Broth) to perform Bacterial cell culture media Escherichia coli

Get tips on using mericon DNA Bacteria Plus Kit (50) to perform DNA isolation / purification Bacteria - Gram positive Clostridium botulinum

Get tips on using SQSTM1/p62 Antibody #5114 to perform Immunohistochemistry Mouse - p62

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.