Kit for completing DNA assembly by depending on T5 exonuclease and PEG8000 and application thereof
A technology of exonuclease and kit, which is applied in the biological field to achieve the effects of low cost, improved efficiency, and convenient operation
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[0064] Example 1. A kit that relies on T5 exonuclease and PEG8000 to complete DNA assembly
[0065] The kit consists of T5 exonuclease and a buffer system containing PEG8000; based on the total system being 20uL, the amount of T5 exonuclease is 0.16U / total reaction system, and the buffer system components of PEG8000 are 115mM, pH=7.5 Tris-HCl, 11mM MgCl 2 , 11mM DTT and PEG8000 with a mass ratio of 6%.
[0066] Among them, the components in the above kits that rely on T5 exonuclease and PEG8000 to complete DNA assembly can be pre-configured to 4 / 3×, 2× or 5× concentration concentration, which is reserved for later addition of DNA, and concentration conditions Does not affect the effect of the assembly reaction. The T5 exonuclease and reaction buffer in the kit are pre-mixed or added later.
[0067] The above kit is tested and stored at -20°C and -80°C without affecting the reaction effect; and the kit can be stored at low temperature for more than half a year without affecting the ...
Example Embodiment
[0068] Example 2. A kit that relies on T5 exonuclease and PEG8000 to complete DNA assembly
[0069] The kit consists of T5 exonuclease and a buffer system containing PEG8000; the kit is based on a total system of 20uL, the amount of T5 exonuclease is 0.04U / total reaction system, and the buffer system component of PEG8000 is 105mM 、Tris-HCl with pH=7.5, 10mM MgCl 2 , 10mM DTT and PEG8000 with a mass ratio of 5%.
[0070] Among them, the components in the above kits that rely on T5 exonuclease and PEG8000 to complete DNA assembly can be pre-configured to 4 / 3×, 2× or 5× concentration concentration, which is reserved for later addition of DNA, and concentration conditions Does not affect the effect of the assembly reaction. The T5 exonuclease and reaction buffer in the kit are pre-mixed or added later.
[0071] The above kit is tested and stored at -20°C and -80°C without affecting the reaction effect; and the kit can be stored at low temperature for more than half a year without affec...
Example Embodiment
[0072] Example 3. A kit that relies on T5 exonuclease and PEG8000 to complete DNA assembly
[0073] The kit consists of T5 exonuclease and a buffer system containing PEG8000; the total system is 20uL, the amount of T5 exonuclease is 0.02U / total reaction system, and the buffer system components of PEG8000 are 110mM, pH=7.5 Tris-HCl, 9mM MgCl 2 , 9mM DTT and 4% PEG8000 by mass.
[0074] Among them, the components in the above kits that rely on T5 exonuclease and PEG8000 to complete DNA assembly can be pre-configured to 4 / 3×, 2× or 5× concentration concentration, which is reserved for later addition of DNA, and concentration conditions Does not affect the effect of the assembly reaction. The T5 exonuclease and reaction buffer in the kit are pre-mixed or added later.
[0075] The above kit is tested and stored at -20°C and -80°C without affecting the reaction effect; and the kit can be stored at low temperature for more than half a year without affecting the reaction effect.
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