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Index / Trevigen / DPO_4+ Buffer + Cations / Product Detail : 4150-050-EB DPO_4+ Buffer + Cations
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Aug
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#4150-050-EB DPO_4+ Buffer + Cations

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  Price : 1074   EUR
1219   USD
833   GBP
4511   Zloty
143712   JPY
8283   NOK
8875   SEK
1214   CHF

Product name : DPO_4+ Buffer + Cations

Catalog number : 4150-050-EB

Quantity: 50 µg

Availability: Yes

Supplier name : Trevigen

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More Details about

Description: Dpo4 is a thermophilic Y-family DNA polymerase capable of performing translesion synthesis and allows for coupling to PCR for amplification of damaged DNA(1). Dpo4 has a molecular weight of 42 kDa and is 95% pure by SDS PAGE. Translesion synthesis is enhanced by the presence of Mn2+ in the reaction.
Source: Purified from E. coli containing a recombinant plasmid harboring the S. solfataricus Dpo4 gene.
Unit Definition: One unit of enzyme incorporates 1 nmole of dTMP onto the 3’ end of d(T)24 using poly(dA)300 as template in 30 minutes at 60°C.
Read-through Specificity: Dpo4 catalyzes translesion synthesis across the following forms of DNA damage: AP sites, cis-syn thymine-thymine dimer, photoproducts, cisplatinum- and acetyl aminofluorene guanine adducts.
Assay Conditions & Analysis: 1X REC Buffer 15 [10 mM HEPES-NaOH (pH 7.8), 1 mM DTT, 50 mM NaCl, 100 µg/ml BSA, 0.1% Triton X-100], 10 mM MgCl2, 1 pmole of 32P-labeled d(T)24, 2 pmole of poly(dA)300, and serial dilutions of enzyme in a 20 µl reaction volume are incubated for 30 minutes at 60°C. Reaction products are resolved by 20% denaturing polyacrylamide gel electrophoresis. Bands are cut out and radioactivity counted to quantify newly synthesized products.
Storage Buffer: 20 mM Tris-HCl (pH 7.8), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) glycerol.
Storage Conditions: Store at –20°C in a manual defrost freezer. For long-term storage, freeze at –80°C in working aliquots. Avoid repeated freeze-thawing. Enzyme may be diluted in 1X REC Buffer 15 for immediate use.

 

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