Enzyme Quality
📄 Certificate of Analysis — COA_Z061_092024_v1
ArtPure™ SP6, free of contaminants.
ArtPure™ SP6 is free of contaminants, ensuring a high yield of pure, properly-capped RNA. Every batch is evaluated across purity, activity, contaminant and in-vivo expression panels.
Purity
RNases
Activity
Proteinases
DNases
Protein expression
Definitions
Activity (U/µl): One enzyme unit incorporates 1 nmol of soluble RNA nucleotides into insoluble polymeric RNA per hour at 37°C.
Purity: Evaluated by SDS-PAGE, where the integrated peak area of the RNA Polymerase is compared to other protein peaks in the same lane.
Transcriptional activity: Quantification of total RNA products after in vitro transcription and RNA purification.
Proteinases: Assessed using Pierce™ Fluorescent Protease Assay Kit (ThermoFisher Scientific) following the manufacturer's guidelines.
RNases: Assessed using the RNase Alert kit (Integrated DNA Technologies), following the manufacturer's guidelines.
Double-stranded endonucleases: 2.5 µl of the SP6 RNAP are incubated with 1 µg of supercoiled plasmid DNA for 0 hours, 4 hours at 37°C and 16 hours at room temperature.
Double-stranded exonucleases: 2.5 µl of the SP6 RNAP are incubated with 0.5 µg of linearized double-stranded DNA substrate for 0 hours, 4 hours at 37°C and 16 hours at room temperature.
Single-stranded exonucleases: 2.5 µl of the SP6 RNAP are incubated with 1 µg of ssDNA substrate (calf-thymus DNA) for 0 hours, 4 hours at 37°C and 16 hours at room temperature.
Protein expression: A linear template containing an SP6 promoter, enhanced Green Fluorescent Protein (eGFP) and poly(A) tail was transcribed and capped using SP6 RNAP and an anti-reverse cap analog (ARCA), injected into zebrafish and analyzed for eGFP expression.