2′-Deoxyuridine 5′-triphosphate (dUTP) may be used to study the distribution, specificity and kinetics of deoxyuridine 5′-triphosphate nucleotidohydrolase(s) (dUTPase, EC 3.6.1.23). dUTPase are important enzymes that protect cells from the incorporation of dUTP into DNA.dUTP is a component of the TUNEL (terminal deoxyribonucleotidyl-transferase-(TdT)-mediated deoxyuridine-5′-triphosphate- (dUTP) nick-end labeling) test. TUNEL is a common method for detecting DNA fragmentation that results from apoptotic signaling cascades.
Applications:Suitable for use in PCR, RT-PCR with Taq DNA Polymerase, cDNA, synthesis and primer extension. Not suitable for PCR with proofreading enzymes or enzyme mixes containing proofreading enzymes. Other applications not tested.
Recommended Dilutions:Optimal dilutions to be determined by the researcher.
Appearance:Supplied as a liquid in 100mM aqueous solution, pH 7.0.
Purity:≥ 99% (HPLC)
Lambda Max:262nm
Endo- and Exonucleases:Incubation of single stranded and double stranded radiolabeled oligonucleotides with 1ul of 20mM dUTP for 4 hours at 37°C, separation on denaturing polyacrylamide gel and phosphoimaging did not detect DNA degradation.
Ribonucleases:Incubation of 2000 base RNA transcript with 1ul of 20mM dUTP at 37°C for 4 hours and separation on agarose gel resulted in no decrease in RNA transcript band intensity compared to control.
Nicking Activities:Incubation of 1ug of supercoiled pUC19 DNA with 1ul of 20mM dUTP at 37°C for 17 hours and separation on agarose gel did not generate linearised plasmid, and relaxation of supercoiled plasmid as compared to control.
E.coli DNA:Quantitative PCR test on ABI Prism 7000 SDS, which uses amplification of E.coli 23S rRNA gene fragment did not detect E.coli DNA.
Human DNA:Quantitative PCR test on ABI Prism 7000 SDS, which uses amplification of human genomic DNA fragment did not detect human DNA.
Functional Test:PCR amplification of human apoprotein gene fragment from 50pg human genomic DNA using Hot Start Taq DNA Polymerase.
Storage and Stability: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.
仕様
Size：100mM
Grade：Molecular Biology Grade
Purity：≥99% (HPLC)
Form：Supplied as a liquid in 100mM aqueous solution, pH 7.0.
