Product Introduction
|
Product Name |
Cat.No. |
Spec. |
|
Taq U DNA Polymerase (for bisulfite-treated DNA) |
G3346-50 |
50 rxns |
Product Description/Introduction
Taq U DNA Polymerase (for bisulfite-treated DNA) is designed to amplify the template DNA containing uracil after bisulfite conversion, which is used for genomic methylation research. The product includes hot-start Taq U DNA Polymerase that can read DNA templates containing uracil. At the same time, the non-specific annealing of primers can be effectively reduced because of the inhibitory effect of antibodies on DNA polymerase at low temperature. 5×Taq U Reaction Buffer is a specially optimized buffer system for amplification of damaged DNA after transformation, which can effectively improve the specificity and yield of amplification.
Storage and Handling Conditions
Shipped with wet ice and stored at -20°C; valid for up to 12 months.
Product Contents
|
Component Number |
Component |
G3346-50 |
|
3346-1 |
Taq U DNA Polymerase |
12.5 µL |
|
3346-2 |
5×Taq U Reaction Buffer |
1.25 mL |
|
3346-3 |
25 mM MgSO4 |
250 µL |
|
3346-4 |
dNTPs (2.5 mM each) |
150 µL |
|
Manual |
One copy |
|
Assay Protocol / Procedures
1. Reaction system (50 μL):
|
Component |
Volume |
|
5×Taq U Reaction Buffer |
10 μL |
|
25 mM MgSO4 |
5 μL |
|
dNTPs (2.5 mM each) |
3 μL |
|
Forward primer (10 µM)a |
2 μL |
|
Reverse primer (10 µM)a |
2 μL |
|
Templateb |
50 ng-100 ng |
|
Taq U DNA Polymerase |
0.25 μL |
|
Nuclease-free Water |
Up to 50 μL |
a: The final concentration of primers ranges from 0.2 to 1.0 μM, and the recommended concentration of primers is 0.4 μM. When there is no amplification product or non-specific amplification, MgSO4 and dNTPs can be adjusted to improve the amplification results.
b: When using the DNA of bisulfite conversion (recommended DNA MethylCode Bisulfite Conversion Kit) as the template, 50-100 ng are recommended for every 50 μL system.
2. Recommend PCR reaction procedure
|
Temperature |
Time |
Cycles |
|
95°C |
10-15 s |
30-40 |
|
55-60°C |
30 s |
|
|
72°C |
1 min/kb |
|
|
72°C |
5-10 min |
1 |
|
4°C |
Forever |
Note
1. It is recommended to use special primer design program to design primers. In order to ensure that the DNA transformed by bisulfite can be well amplified, it is recommended that the amplified fragments should not exceed 500 bp when designing primers.
2. The optimization of PCR reaction can be adjusted from the aspects of Mg2+ concentration, template amount, enzyme amount, annealing temperature and so on.
3. Repeated freeze-thaw DNA template will affect the amplification, if you need many experiments, can be sub-packed and frozen to reduce the number of freeze-thaw.
For Research Use Only!
