Protein Expression Eukaryotic cells K. lactis

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Get tips on using Gibco™ DMEM, high glucose, GlutaMAX™ Supplement to perform 3D Cell Culture Media Murine colonic organoids

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Get tips on using Rock-2 siRNA and shRNA Plasmids (h) to perform RNA sequencing Human - HT-1376 (urinary bladder cell line)

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Get tips on using DMEM/HAM'S F-12 LIQUID MEDIUM WITHOUT L-GLUTAMINE to perform Mammalian cell culture media RBMVEC

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Get tips on using Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham to perform Mammalian cell culture media DU145

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Get tips on using Dulbecco’s Modified Eagle’s Medium/Nutrient Mixture F-12 Ham to perform Mammalian cell culture media T47D

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Get tips on using IncuCyte® Caspase-3/7 Apoptosis Assay Reagent to perform Apoptosis assay cell type - Caspase 3/7

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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.

RNA cDNA synthesis Cell lines

Get tips on using DMEM, no glucose, no glutamine, no phenol red to perform 3D Cell Culture Media hiPSC-derived cardiac organoids

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Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform 3D Cell Culture Media hiPSC-derived retinal organoids

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Get tips on using Gibco™ DMEM/F-12, GlutaMAX™ supplement to perform 3D Cell Culture Media hESC-derived brain organoids

Products Thermo Fisher Scientific Gibco™ DMEM/F-12, GlutaMAX™ supplement

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