2’-Deoxyuridine 5’-Triphosphate, Sodium Salt (dUTP) Solution

Blue Ice

534, 1

Supplied as a liquid in 100mM aqueous solution, pH 7.0.

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.