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Rapid and Economical Cloning |
Description
RBC T&A cloning system is ideal for cloning PCR products generated using thermostable DNA polymerases which add a single terminal 3'-dA nucleotide overhang (Taq). It is extremely economical and convenient for rapid cloning. Following ligation, the mixture may be used directly to transform HIT Competent Cells™ or other competent cells or purified to achieve a higher efficiency of or other competent cells or purified to achieve a higher efficiency of transformation. The vector is highly purified to reduce background cloning. |
Applications
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- aCCEPTS TERMINAL 3'-dA nucleotide overhang PCR products.
- Transform ligation product (purified/unpurified) into HIT Competent Cells™.
- LacZ complementation for blue/white screening.
- Ampicillin marker for antibiotic selection.
- Universal primer for easy transformation screening.
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Cat. No. RC001 (20 reactions)
TA Cloning Vector (25 ng/μl): 40 μl
Control Insert DNA (10 ng/μl): 10 μl
T4 DNA Ligase (3 U/μl): 20 μl
T4 DNA Ligation Buffer A: 100 μl
T4 DNA Ligation Buffer B: 100 μl
Forward Primer(M13-F) (10 μM): 50 μl
Forward Primer(M13-R) (10 μM): 50 μl |
Ligation Conditions |
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User Sample |
Positive Control |
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| Ligation Buffer A |
1 μl |
1 μl |
| Ligation Buffer B |
1 μl |
1 μl |
| TA cloning vector |
2 μl |
2 μl |
| PCR product |
X μl |
**** |
| T4 DNA Ligase |
1 μl |
1 μl |
| Control DNA |
**** |
3 μl |
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Add deionized water to a final volume of 10 μl. Incubate the reactions for 5 to 15 mins at 22°C
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Storage Conditions |
T&A Cloning Kit should be stored immediately upon receipt at -20°C in a constant temperature freezer. T&A Cloning Kit can be stored for up to 12 months without showing any deduction in performance and quality with proper storage.
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Note: T&A is a rapid high quality cloning system-Following are comparison results from RBC Labs. |
- TA cloning system is very efficient at LIGATION with higher resulting transformation effciency compared to company P's transformation system. T&A cloning system provides two types of ligation buffer for your convenience.
- T&A cloning system high efficiency ligation takes only 20 minutes versus 1 hour to overnight for company P's system.
- TA cloning system resulted in higher ACCURACY compared to company P's in colony PCR pickings.
- TA cloning system shows very good effciency of cloning up to 5 kb INSERTS, while company P's system may show slightly higher effciency > 5kb.
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