Alw21I (HgiAI)

Produktübersicht

• Artikelname: Alw21I (HgiAI)
• Artikelnummer: USB-A1372-05
• Hersteller Artikelnummer: A1372-05
• Alternativnummer: USB-A1372-05-500
• Hersteller: US Biological
• Kategorie: Molekularbiologie

Produktbeschreibung

5 - G W G C W C - 3 3 - C W C G W G - 5 Source: Acinetobacter lwoffi RFL21 Concentration: 10u/ul Form: Supplied as a liquid in 10mM Tris-HCl pH 7.5, 300mM sodium chloride, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Dilution Buffer: 10mM Tris-HCl pH 7.4, 100mM potassium chloride, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol. Unit Definition: One unit is defined as the amount of Alw21I required to digest 1ug of lambda DNA in 1 hour at 37C in 50ul of assay buffer. Supplied With: R1625: Restriction Enzyme Buffer A, 10X: Supplied as a liquid in 33mM Tris-acetate, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA pH 7.9 at 37C when diluted to 1X. Note: 1X conc. yields 20-50% enyzme activity. 2X conc. yields 50-100% enzyme activity. R1625-03: Restriction Enzyme Buffer D, 10X: Supplied as a liquid in 50mM Tris-HCl, pH 7.5, 10mM MgCl2, 100mM sodium chloride, 0.1mg/ml BSA when diluted to 1X. Incubate at 37C. Note: This buffer yields 100% enzyme activity Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with Alw21I. Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with Alw21I, more than 95% of the DNA fragments can be ligated at a 5-termini concentration of 0.27uM. More than 95% of these sites can be recut. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of Alw21I for 4 hours. Stability During Prolonged Incubation: A minimum of 0.1 units of Alw21I is required for complete digestion of 1ug of lambda DNA in 16 hours at 37C. Compatible Ends: GTGCA C - BseSI, SduI, Mph1103I, PstI, SdaI GAGCT C - Eco24I, SacI, SduI GAGCA C - SduI Number of Recognition Sites in DNA: Lambda: 28 PhiX174: 3 M13mp18/19: 3 pBR322: 8 pUC18/19: 5 pUC57: 5 pTZ19R/U: 4 Thermal Inactivation: Alw21I is inactivated by incubation at 65C for 20 min. Recommended Protocol for Digestion: Add: Nuclease free water: 16ul 10X Restriction Enzyme Buffer D: 2ul DNA (0.5-1ug/ul): 1ul Alw21I: 0.5-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. This digestion reaction may be scaled either up or down. Protocol for Digestion of PCR Products Directly After Amplification: Add: PCR Reaction Mixture: 10ul (~0.1-0.5ug of DNA) Nuclease free water: 18ul 10X Restriction Enzyme Buffer D: 2ul Alw21I: 1-2ul Mix gently, spin down for a few seconds. Incubate at 37C for 1-16 hours. Methylation Effects on Digestion: Dam, Dcm: Never overlaps, no effect CpG, EcoKI: May overlap, no effect EcoBI: May overlap, effect not determined Storage and Stability: May be stored at 4C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20C. Aliquots are stable for 6 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.