P3121
PdiI (NaeI)
Size
£269.00
- SKU:
- P3121
- Storage Conditions:
- -20°C
- Supplier:
- United States Biological
- Extra Details:
- 5'-G C C^G G C-3' 3'-C G G^C C G-5' Concentration: 10u/ul Source: Pseudomonas diminuta Mck 33–321 Stability during Prolonged Incubation: A minimum of 0.5 units of P3121 is required for complete digestion of 1ug of DNA in 16 hours at 37°C. Thermal Inactivation: Enzyme is inactivated by incubation at 65°C for 20 minutes. Digestion of Agarose-embedded DNA: A minimum of 20 units of P3121 is required for complete digestion of 1ug of agarose- embedded pBR322 DNA in 16 hours. Unit Definition: One unit is defined as the amount of P3121 required to digest 1ug of pBR322 DNA-NdeI fragments in 1 hour at 37°C in 50ul of reaction bu Number of Recognition Sites in DNA: Lambda: 1 PhiX174: 0 pBR322: 4 pUC18/19: 0 pUC57: 0 pTZ19R/U: 1 M13mp18/19: 1 Recommended Protocol: Add: Nuclease-free water 16ul R1625: 2ul DNA: 1ul P3121: 0.5-2ul 1. Mix gently and spin down for a few seconds. 2. Incubate at 37ºC for 1-16 hours. Protocol for Digestion of PCR after Amplification: Add: PCR mixture: 10ul Nuclease-free water: 18ul R1625: 2ul P3121: 1-2ul 1. Mix gently and spin down for a few seconds. 2. Incubate at 37ºC for 1-16 hours. Menthylation Effects on Digestion: No effect on Dam, Dcm, EcoKl and EcoBl. Overlaps and blocks CpG. Supplied with: R1625: Restriction Enzyme Buffer A, 10X: Dilute to 1X for use. 1X buffer composition is 33mM Tris-acetate pH 7.9 at 37°C, 10mM magnesium acetate, 66mM potassium acetate, 0.1mg/ml BSA. Diluent Buffer: 10mM Tris-HCl, pH, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. Storage Buffer: Supplied as a liquid in 10mM Tris-HCl, pH 7.4, 500mM KCl, 1mM DTT, 0.1mM EDTA, 0.2mg/ml BSA, 0.15% Triton X-100, 50% glycerol. Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 160-fold overdigestion (10u/ug lambda DNA x 16 hours) with P3121. Ligation and Recleavage (L/R) Assay: The ligation and recleavage assay was replaced with L0 test after validating experiments showed L0 test abililty to trace nuclease and phophatase activities with senstivity that is higher than L/R by a fctor of 100. Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide occurred during incubation with 10 units of Pdil for 4 hours. Blue/White Cloning Assay: The B/W assay was replaced with L0 test after validating experiments showed L0 test ability to detect nuclease and phophatase activities with sensitivity that equals to that of B/W test. Methylation Effects: No effect on Dam, Dcm, EcoKl and EcoBl. Overlaps and blocks CpG. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
- Shipping Conditions:
- Blue Ice