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 effciency of or other competent cells or purified to achieve a higher effciency of transformation. The vector is highly purified to reduce background cloning.
Applications
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.
Ligation Conditions
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User Sample
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Positive Control
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Ligation Buffer A
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1 μl
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1 μl
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Ligation Buffer B
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1 μl
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1 μl
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T&A cloning vector
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2 μl
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2 μl
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PCR product
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X μl
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****
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T4 DNA Ligase
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1 μl
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1 μl
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Control DNA
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****
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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.
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.
Cat. No. RC001 (20 reactions)
T&A 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
Cat. No. RC013 (20 reactions)
T&A Cloning Vector (25 ng/μl): 40 μl
Note: T&A is a rapid high quality cloning system-Following are comparison results from RBC Labs.
| i) |
T&A cloning system is very effcient 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. |
| ii) |
T&A cloning system high effciency ligation takes only 20 minutes versus 1 hour to overnight for company P's system. |
| iii) |
T&A clonng system resulted in higher accuracy compared to company P's in colony PCR pickings. |
| iv) |
T&A 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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