Protein Expression Prokaryotic cells E. coli

Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - Point mutation Neuro 2a Epac1

Get tips on using mirVana™ miRNA Isolation Kit, with phenol to perform RNA isolation / purification Bacteria - Gram negative Salmonella enterica

Get tips on using ON-TARGETplus Mouse Hspa5 (14828) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 Grp78/Hspa5

Get tips on using ON-TARGETplus Mouse Mapk14 (26416) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 p38/Mapk14

Get tips on using NucleoSpin® Gel and PCR Clean-up to perform DNA gel extraction / PCR product purification Product size > 15Kb

Get tips on using MagNA Pure Compact Nucleic Acid Isolation Kit I to perform DNA isolation / purification Bacteria - Gram negative Enterobacteriaceae

Get tips on using PowerSoil® DNA isolation - DNeasy PowerSoil Pro Kit to perform DNA isolation / purification Bacteria - Gram negative Enterobacteriaceae

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 MagNA Pure Compact Nucleic Acid Isolation Kit I to perform DNA isolation / purification Bacteria - Gram negative Salmonella enterica