SP6 RNA Polymerase

English name：SP6 RNA Polymerase
Other name: SP6 RNA polymerase

Grade：BR
Molecular weight：The monomer is about 98kDa

Source: E. coli cells containing cloned genes encoding corresponding RNA polymerase.

Concentration：20u/ul（≥100U/ul，HC）
Activity definition: Refers to the amount of enzyme required to synthesize and incorporate 1nmol amp into oligonucleotide fragment (DE-81) light absorption form) within 60 minutes at 37 ℃.

Enzyme activity analysis mixture：40mM Tris-HCL(PH8.0), 10mM DTT, 6mM MgCL2, 2mM Spermidine，0.5mM Various NTP, 0.6MBq/ML(3H).-ATP和20ug/ml:  Plasmid DNA containing corresponding promoter sequence.

Save buffer：Each polymerase is stored in  50mM Tris-HCL(PH8.0), 5mM DTT, 0.1mg/ml BSA,150mM NaC1, 0.5mM ELUGENT  Detergent  and  50%(V/V) glycerol.

5× Transcription buffer：200mM Tris -HCL(PH9.0,25℃), 30mM MgCL2, 50mM DTT, 50mM NaC1和10mM Spermidine.

Inhibitor：Metal chelating agent，When the concentration of NaC1 or KC1 is higher than 150 mm (SP6 and T7 RNA polymerase), the enzyme activity decreases by more than 50%, and ammonium sulfate can reduce the enzyme activity by more than 50%.

Deactivation: Add EDTA or heat at 70 ℃ for 10 minutes.

Quality control: Relevant tests show that there is no internal or external deoxyribonuclease and ribonuclease pollution

Functional test: In vitro transcription reaction

Features: synthesis of nucleotides mixed with special nucleotides, such as amino -, biotin -, fluorescein -, digoxin labeled nucleotides

Physicochemical properties: liquid SP6 RNA polymerase is a DNA dependent polymerase with high specificity for SP6 promoter sequence.When RNA is synthesized with this enzyme, only SP6 DNA or DNA cloned downstream of SP6 can be used as the synthesis template.The enzyme can take 32P, 35S and 3H labeled nucleoside triphosphate as substrate.

Purpose: Biochemical research.Synthesis of labeled and unlabeled RNA for hybridization and in vitro RNA translation;As a substrate for RNA, siRNA and RNase digestion protection analysis and a template for genomic DNA sequencing;Study the secondary structure of RNA, RNA protein interaction and RNA splicing; Labeled RNA probes were synthesized for Western blot, in situ hybridization and microarray hybridization.