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JoVE Journal

Bioengineering

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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

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08:31 min

February 5th, 2021

February 5th, 2021

12,543 Views

0:04

Introduction

0:26

Cloning Parts into the Entry Vector (pYTK001) to Create Part

2:05

Assembling Part Plasmids into Cassette Plasmids

4:50

Assembling Cassette Plasmids into Multi-Gene Plasmids

5:55

Results: Multi-gene Plasmid Cloning in E. coli and Yeast

7:54

Conclusion

Transcrição

This is a detailed protocol for assembling multi-gene plasmids using the Yeast MoClo kit. This method enables the convenient cloning of a variety of multi-gene classmates. It is ideal for generating a big library of related parts.

Begin by s

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The goal of this protocol is to provide a detailed, step-by-step guide for assembling multi-gene constructs using the modular cloning system based on Golden Gate cloning. It also gives recommendations on critical steps to ensure optimal assembly based on our experiences.

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