T4 Ligase
E. coli lambda lysogen NM 989
| Catálogo Nº | Apresentação | Preço (R$) | Comprar |
|---|---|---|---|
| ENZ-101S | 100 U | 270,82 | Adicionar ao Carrinho |
| ENZ-101M | 250 U | 431,34 | Adicionar ao Carrinho |
| ENZ-101L | 2 x 250 U | 718,54 | Adicionar ao Carrinho |
| ENZ-101XL | 4 x 250 U | 1.179,36 | Adicionar ao Carrinho |
For in vitro use only!
Definição de unidade: One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmol of 32P from pyrophosphate to ATP, into Norit-adsorbable material in 20 minutes at 37 °C.
Envio: shipped on gel packs
Condições de armazenamento: store at -20 °C
avoid freeze/thaw cycles
Validade: 12 months
Forma: liquid (Supplied in 10 mM Tris-HCl pH 7.4, 50 mM KCl, 0.1 mM EDTA, 1 mM DTT, 200 μg/ml BSA and 50 % [v/v] glycerol)
Concentração: 2.5 Weiss units/μl (500 CE units/μl)
Descrição:
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5’ phosphate and 3’-hydroxyl termini in duplex DNA or RNA.
Contente:
Standard Ligation Buffer, 10x conc.
500 mM Tris-HCl pH 7.8 at 25 °C, 100 mM MgCl2, 100 mM DTT, 10 mM ATP and 25 μg/ml BSA
Fast Ligation Buffer, 2x conc.
60 mM Tris-HCl pH 7.8 at 25 °C, 20 mM MgCl2, 20 mM DTT, 2 mM ATP and 10 % PEG
Heat inactivation:
T4 DNA Ligase can be inactivated by incubation at 65 °C for 10 minutes.
Note:
• One Cohesive-End Ligation Unit (CEU) is defined as the amount of enzyme required to give 50 % ligation of Hind III fragments of λ DNA (5’ DNA termini concentration of 0.12 μM, 300 μg/ml) in a total reaction volume of 20 μl in 30 minutes at 16 °C in 1x T4 DNA Ligase Reaction Buffer.
• One Weiss unit is equivalent to approx. 200 CE units.
• T4 DNA Ligase is strongly inhibited by NaCl or KCl if the concen- tration exceeds 200 mM.
• Ligation of blunt-ended and single-base pair overhang frag- ments requires about 50 times as much enzyme to achieve the same extent of ligation as cohesive-end DNA fragments. Blunt- end ligation may be enhanced by addition of PEG 4000 (10 % w/v final concentration) or hexamine chloride, or by reducing the ATP concentration to 50 μM.
• To dilute T4 DNA Ligase for subsequent storage at -20 °C a storage buffer containing 50 % glycerol should be used, to dilute Ligase for immediate use, 1x Reaction Buffer is recommended.