Pro Ligation-Free Cloning MasterMix
Cat. No.
G4004
Unit
50 Reactions
Price
$258.00
| Cat. No. | G4004 | |||||||||||
| Name | Pro Ligation-Free Cloning MasterMix | |||||||||||
| Unit | 50 Reactions | |||||||||||
| Category | Molecular Biology Enzymes and Kits | |||||||||||
| Description |
abm’s Pro Ligation-Free Cloning MasterMix is a ready-to-use solution designed for rapid, efficient and seamless DNA assembly regardless of fragment length or end compatibility. It enables one-step assembly of multiple DNA fragments with overlapping ends in a single isothermal reaction making it ideal for synthetic biology and molecular biology applications. To support accurate fragment amplification prior to assembly, the kit includes MegaFi™ Pro Fidelity 2X PCR MasterMix to enable robust, high-fidelity PCR. Key Features:
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| Application |
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| Storage Condition | Store all components at -20°C | |||||||||||
| Note |
Retired Cat. No. E086 and E087 |
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G4004 |
Datasheet
Search CoA here
MSDS
| How many fragments of DNA can be assembled in one reaction? | |
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The technology is capable of assembling >10 fragments in a single reaction according to the literature. In house experiments have routinely achieved 6 fragment assemblies.
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| What are the shortest overlaps that can be used with this assembly method? | |
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abm recommends a minimum of 15 bp of overlapping sequence with a Tm >50°C. Increasing the length of the overlap can also increase the efficiency of the assembly reaction.
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| How do I optimize conditions for 3+ fragment assembly? | |
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abm recommends the following:
1. Setting up the reaction where all components (vector and inserts) are added in equimolar amounts.
2. Increasing the incubation time to 60 min.
3. Utilizing high transformation efficiency electrocompetent E. coli cells.
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| How do I design overlapping primers for more than one fragment? | |
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You can use the same logic outlined in the product Datasheet Appendix. See the below image for a multi-fragment assembly example:
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| Do I need to gel extract and/or purify my DNA fragments before assembly? | |
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This is an optional but recommended step. Gel purification improves efficiency and accuracy of the DNA assembly especially for complex, multi-fragment assemblies as it removes non-specific PCR products, primer-dimers etc.
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| After the assembly product has finished incubation, do I need to transform it into competent cells? | |
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Yes, this is a necessary step that cannot be skipped in the DNA assembly protocol.
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