2×Taq PCR Super Fast Mix(with blue dye) |
| Catalog No.GK10035 |
2×Taq PCR Super Fast Mix(with blue dye) is a ready-to-use, rapid, and simple pre-mixed solution for PCR that contains a blue loading buffer, and it is suited for super-fast PCR reactions.
Products are for research use only. Not for human use. We do not sell to patients.
Sample solution is provided at 25 µL, 10mM.
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2×Taq PCR Super Fast Mix(with blue dye) is a ready-to-use, rapid, and simple pre-mixed solution for PCR that contains a blue loading buffer, and it is suited for super-fast PCR reactions.
The reaction mixture only needs to add primers, template, and ddH2O. 2×Taq PCR Master Mix contains blue loading buffer already, which saved the step of mix loading buffer, and could indicate the electrophoresis progress after the PCR reaction. The migration speed of the blue dye in 1% TAE agarose gel is similar to a 500 bp double-stranded DNA fragment.
2×Taq PCR Super Fast Mix(with blue dye) is well-suited for super-fast PCR reactions, achieving an extension speed of 5-10 sec/kb, enabling the completion of the shortest PCR in as little as half an hour. It efficiently tackles complex templates with high GC content ( >60% ) and secondary structures, while also supporting large-scale gene detection. When working with short fragments or simple templates, such as plasmid templates, using an extension speed of 5 sec/kb can help reduce cycles and further shorten the PCR time, for long fragments ( ≥ 3 kb ) or complex templates producing low yields, an extension speed of 15-20 sec/kb or an increase in cycles may be required.
This product is suitable for large-scale genetic testing, semi-quantitative PCR experiments, and detection of trace DNA; it also facilitates rapid PCR amplification for cloning of expressed genes, analysis of intracellular gene point mutations (SNP), and other purposes.
Application Recommendations:
This product has a very strong amplification ability, and sometimes non-specific amplification may occur. The following methods can be used to reduce non-specific amplification bands.
(1) Increase the annealing temperature by 2-3 degrees;
(2) Reduce the mix volume, such as changing 10 μl to 8 μl;
(3) Reduce the number of cycles by 2 to reduce nonspecific amplification bands.
This protocol only provides a guideline, and should be modified according to your specific needs.
Note: Template dosage: 10-1000 ng genomic DNA; 1 to 30 ng of plasmid or 1 to 2μl of cDNA after RT-PCR reaction. The above example is a conventional PCR reaction system, which is for reference only. The actual reaction conditions vary with different structures such as templates and primers. The best reaction conditions should be set according to the characteristics of templates, primers and target fragments, and the reaction system should be enlarged or reduced according to the proportion.
Frequently Asked Questions (FAQs):
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Common problem |
Possible Causes |
Solutions |
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Low or no PCR product. |
Inappropriate primer design. |
Choose appropriate primer design software for primer design. |
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To obtain high GC content or long fragments. |
Use more suitable DNA polymerase such as 2×Taq PCR Fusion-Fast Mix (with blue dye) (Catalog: GK10036). |
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The annealing temperature is not appropriate; The extension period is short; The number of cycles is insufficient. |
Optimizing PCR experimental conditions. |
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Low template concentrations; experimental sample is damaged or degraded. |
Increase the template volume appropriately. Reprepare templates. |
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The quality of the template is poor. |
Purify template DNA by appropriate DNA purification methods (e.g. column purification). |
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Inappropriate primer concentration and Mg2+ concentration. |
Adjust primer concentration and Mg2+ concentration appropriately. |
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Amplification of nonspecific products in quantitative polymerase chain reactions (qPCR) |
The primers showed nonspecific binding and dimerization. |
Reduce the primer concentration and re-design the primer if necessary. |
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The quality of the template is poor ; templates contaminated or degraded. |
Check template integrity and concentration by Gel electrophoresis and repurify template if necessary. |
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Inappropriate cycling conditions. |
If there are stray bands smaller than the target band, it can be adjusted by increasing the annealing temperature and lowering the number of cycles; if there are stray bands larger than the target band, shorten the elongation time and lower the number of cycles. |
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Amplification products appear in the blank control |
Inappropriate primer design. |
Amplified sequences are homologous to non-target amplified sequences, and PCR can also amplify sequences that are not target sequences, redesigning primers if necessary. |
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PCR reaction system was contaminated. |
If the amplification product band size is not consistent with the target band, it indicates contamination; Repeat the experiment by replacing the Mix, water or primer with a new one; Prepare the reaction system in an ultra-clean bench to reduce aerosol pollution. |
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No Band or Faint Band |
Primer degradation. |
Check primers for degradation due to improper storage conditions and replace primers if necessary. |
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Poor Template quality. |
Template quality should be confirmed before PCR reaction. Improper long-term storage and repeated freezing and thawing may cause template breakage , loop opening or degradation, and prepare DNA template freshly ; if the templates are no longer usable, the products of the first amplification can be diluted in multiplicity to be used as templates for the second amplification. |
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Enzyme concentration. |
It is recommended to increase the amount of enzyme. |
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Inappropriate cycling conditions. |
Perform the Touch Down program ; Optimize the Extension Time and set more cycles. |
| Applications |
1. Genetic testing: The error between different batches of this product is very small, and it is particularly suitable for large-scale genetic testing, semi-quantitative PCR experiments and detection of trace DNA.
2. Ultrafast PCR amplification: such as cloning of expressed genes, analysis of intracellular gene point mutations (SNP), etc. 3. The PCR product has a 3' end dA overhang, which can be used for subsequent TA cloning. |
| Shipping | Ship with blue ice. |
| Storage Conditions | For long-term storage, please store at -20˚C, valid for 24 months. For frequent use, store at 4˚C for at least six months. |
| Usage | For research use only! Not for use on humans. |
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
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