Cat.No.：A03021A
Price: ￥460
Specifications：
This product has been optimized based on the seamless cloning kit, with the addition of stabilizers, which has enhanced the seamless cloning efficiency for long and multiple fragments.
Efficient connection of multiple and long fragments: For seamless cloning of multiple fragments, it can be connected to up to 10 DNA fragments with a total length of 8 kbp or 4 DNA fragments with a total length of 10 kbp; for seamless cloning of long fragments, it can connect a single DNA fragment up to 10 kbp in length.
Product stability is enhanced: After repeated freeze-thawing 20 times, approximately 50% of the activity can still be retained.
| Cat. No. | Component | Specification | Quantity |
| B03021A | PLUS Seamless Cloning Mix (2×) | 100 μL | 1 |
| B03022A | PLUS Seamless Cloning Mix (2×) | 200 μL | 1 |
Store at -20℃ and transport at ≤0℃.
This product can connect up to 10 DNA fragments with a total length of 8 kbp or 4 DNA fragments with a total length of 10 kbp to a linearized vector through homologous recombination in a single reaction.
How to choose the seamless cloning kit and the enhanced seamless cloning kit?
The seamless cloning kit can meet the seamless cloning needs in most scenarios. It can achieve seamless cloning for up to 4 fragments with a total length of 4000bp when dealing with multiple fragments. If the number of used fragments is greater than 4, or the length of a single fragment is greater than 4000bp, it is recommended to use the enhanced seamless cloning kit.
When using the enhanced seamless cloning kit, are there any restrictions on the number and size of the inserted fragments?
The total number of fragments should be no more than 8 and the total length should be no more than 10,000 base pairs. There is no size limit for individual fragments.
When performing multi-segment seamless cloning, there are a large number of false positive transformants with self-circle formation of vectors or incomplete recombination. How to solve this problem?
For multi-segment seamless cloning, it is advisable to appropriately extend the homologous arms between the fragments (e.g. 35bp - 50bp) to increase the correct recombination rate. Additionally, it is necessary to ensure that the vector used is completely linearized.
When performing seamless cloning of long fragments, a large number of false positive transformants are produced. How to solve this problem?
Whether it is single-fragment seamless cloning or multi-fragment seamless cloning, if the length of a single fragment is ≥ 4000 bp, it is necessary to consider reducing the total amount of nucleic acids in the seamless cloning reaction system to below 100 ng.
When performing multi-segment seamless cloning, the number of transformants is relatively small.
The number of transformation vectors is inversely proportional to the number of insertion fragments and the total length. It is advisable to use Zymagen5α competent cells for the transformation experiment, which can significantly increase the number of transformants.
