BstEII

Isoschizomers: BstPI, Eco91I, EcoO65I, PspEI

Catálogo Nº Apresentação Preço (R$) Comprar
EN-144S 1.750 UnitsSob demanda Adicionar ao Carrinho
EN-144L 5 x 1750 UnitsSob demanda Adicionar ao Carrinho
5'–GGTNAC C–3'
3'–C CANTGG–5'

For general laboratory use.

Definição de unidade: One unit is the amount of enzyme required to completely digest 1 μg of Lambda DNA (13 sites) in 1 hour in a total reaction volume of 50 μl. Enzyme activity was determined in the recommended reaction buffer.

Envio: shipped on gel packs

Condições de armazenamento: store at -20 °C
avoid freeze/thaw cycles

Validade: 12 months

Forma: liquid (Supplied in 10 mM Tris-HCl pH 7.4, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA and 50 % [v/v] glycerol)

Concentração: 10 units/μl

Source: Bacillus stearothermophilus Supplied with: 10x Universal Buffer (UB) Recommended 50 μl assay5 μl||10x Universal Buffer (UB) 1 μg||pure DNA1 or PCR product2 5 units||enzyme fill up to 50 μl||PCR grade water 1 Supercoiled or high molecular weight DNA (e.g. plant genomic DNA) may require longer incubation time or higher amount of enzyme. 2 Some enzymes may require additional DNA bases flanking the restriction site for complete digestion. Protocol:The enzyme should not exceed 10 % of total reaction volume.Add enzyme as last component. Mix components well before adding enzyme. After enzyme addition, mix gently by pipetting. Do not vortex.Incubate 60 min. at 60 °C.Stop reaction by alternatively: - Addition of 2.1 μl EDTA pH 8.0 [0.5 M], final 20 mM - Heat Inactivation (No) - Spin Column DNA Purification (e.g. PCR Purification Kit, Cat.-No. PP-201S/L) - Gel Electrophoresis and Single Band Excision (e.g. Agarose Gel Extraction Kit, Cat.-No. PP-202S/L) - Phenol-Chloroform Extraction or Ethanol Precipitation. Double Digestion - Buffer Compatibility: B1 - 50 % Relative Activity B2 - 50-75 % Relative Activity B3 - 75-100 % Relative Activity B4 - 50 % Relative Activity B5 - 75 % Relative Activity 1x UB - 100 % Relative Activity (recommended) Please note that the optimum digestion condition for this enzyme is 1x UB. Within the Universal Buffer (UB) system, the most majority of our enzymes display 100% Relative Activity in 1x UB and only few either in 0.5x or 2x UB. If optimum condition for second enzyme is different than the recommended for the first enzyme, we suggest carrying out first the restriction at the higher recommended concentration of UB and dilute the reaction volume to the adequate UB concentration for further proceeding with the second restriction. Reaction Enzymes Buffer Guide:Buffer 1||10 × B1||100 mM 100 mM1000 μg/ml||Tris-HCl(pH 7.9, 25°C)MgCl2BSA Buffer 2||10 × B2||100 mM 100 mM500 mM1000 μg/ml||Tris-HCl(pH 7.9, 25°C)MgCl2NaClBSA Buffer 3||10 × B3||500 mM 100 mM1000 mM1000 μg/ml||Tris-HCl(pH 7.9, 25°C)MgCl2NaClBSA Buffer 4||10 × B4||100 mM 100 mM1500 mM1000 μg/ml||Tris-HCl(pH 7.9, 25°C)MgCl2NaClBSA Buffer 5||10 × B5||200 mM 100 mM500 mM1000 μg/ml||Tris-acetate(pH 7.9, 25°C)Mg-acetateK-acetateBSA Reaction Buffer Compatibility: Our restriction enzymes are fully compatible to restrictases and buffer systems from other manufacturers and can be used along in double digestions. To obtain best results, consult the corresponding manuals of all involved products. Ligation and recutting: After 40-fold overdigestion with BstEII, >95 % of the DNA fragments can be ligated and recut with this enzyme. Note: BstEII exhibits 10-15 % activity at 37 °C. DNA Methylation: No Inhibition: dam, dcm, CpG Quality Control: All preparations are assayed for contaminating endonuclease, 3'-exonuclease, 5' exonuclease/ 5' phosphatase, as well as nonspecific single- and doublestranded DNase activities.

Produtos relacionados: Universal Restriction Buffer, #EN-300