Takara In-Fusion Snap Assembly Starter Bundle \10 Rxns

In-Fusion Snap Assembly Master Mix enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes:

- A NucleoSpin Gel and PCR Clean-Up kit: This kit is suitable for gel extraction as well as PCR purification. Kits are provided with individual purification columns.

- Stellar Competent Cells: High-efficiency competent cells are essential to the success of In-Fusion Cloning. AnE. coliHST08 strain is included that provides high transformation efficiency (greater than 5 x 10^8 CFU/µg) and complements the efficiency of all In-Fusion Snap Assembly kits. Cells are provided in 100-μl aliquots in individual tubes.

- PrimeSTAR Max DNA Polymerase: This convenient 2X liquid master mix offers exceptionally accurate, efficient, and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid setup of PCR reactions, and facilitates successful cloning. The polymerase master mix is provided in 625-μl aliquots.

Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.

The In-Fusion cloning protocol.