Products
Recombinant Protein & Enzyme
Modification Enzyme
DNA/RNA Polymerase
Isothermal Amplification and Ligase Chain Reaction Enzymes
SKU : G3432-250U
Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat.No. | Spec.. |
Bsu DNA polymerase, large Fragment | G3432-250U | 250 U |
Product Description/Introduction
Bsu DNA polymerase LargeFragment is from bacillus subtilis DNA polymerase, larger pieces of E. coli recombinant expression, 5'-3' DNA polymerase activity, do not have 3'-5' and 5'-3' exonuclease activity. Bsu DNA polymerase, LargeFragment also have chain replacement activity, are often used to restructure polymerase enzyme amplification (RPA) reaction, the temperature of the isothermal amplification is usually 37.
Uses: Isothermal amplification by RPA method; RPA strand replacement DNA synthesis; Random primer labeling; cDNA second strand synthesis.
Source: Derived from Bacillus subtilis, recombinant expression by Escherichia coli.
Inactivation or inhibition: complete inactivation for 20 minutes at 75.
Enzyme storage buffer:25 mM Tris-HCl，50 mM NaCl，1 mM DTT，0.1 mM EDTA，50%Glycerol，pH 7.4.
10×Bsu Reaction Buffer：100 mM Tris-HCl，500 mM NaCl，100 mM MgCl2，10 mM DTT，pH 7.9.
Storage and Shipping Conditions
Wet ice transport; Store at -20 for 12 months.
Product Content
Component Number | Component | G3432-250U |
G3432-1 | Bsu DNA polymerase, large Fragment | 50 μL |
G3432-2 | 10×Bsu Reaction Buffer | 500 μL |
Manual | One copy | |
Assay Protocol / Procedures
Examples of isothermal amplification:
1. Single-strand primer and DNA Template were mixed with equal molar number, the recommended final concentration was 10μM, incubated at 95 for 2min, and then gradient cooling to 25 annealing to form Primer/Template hybrid double-strand.
2. Prepare the reaction system according to the following table:
Component | Volume |
Nuclease-Free Water | 15 μL |
Primer/Template | 1 μL |
dNTP Mix（2.5 mM each） | 1 μL |
10×Bsu Reaction Buffer | 2 μL |
Bsu DNA polymerase, large Fragment | 1 μL |
3. Incubate the above reaction system at 37 for an appropriate time (the incubation time needs to be explored in actual use). The elongation rate tested was similar to that of conventional DNA polymerase, about 1kb/min.
4. After incubation at 75 for 20min to stop the reaction, agarose gel electrophoresis can be performed.
Note
1. Enzyme products should be placed in an ice box or ice bath when used, and should be stored at -20 immediately after use, it is recommended to store in separate packages.
2. Bsu DNA polymerase largeFragment because of the lack of 3'-5' exonuclease activity, should not be used for the production of flat end.
3. Bsu DNA polymerase, a large Fragment, retains about 50% of its activity at 25 twice as much as the Klenow fragment (3'-5' exo-) at this temperature.
4. For your safty and health,please wear safety glasses, gloves, or protective clothing.
For Research Use Only!
