A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing
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11:36 min
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July 3rd, 2016
DOI :
July 3rd, 2016
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0:05
Title
0:43
Synthesis of NNS Sub-libraries for Each Amino Acid Position by Two-step PCR Site-directed Mutagenesis
1:53
Cloning of NNS Sub-libraries into Selection Vectors
3:47
Selection of the TEM-1 Whole-Protein Saturation Mutagenesis Library for Antibiotic Resistance
5:51
High Throughput Sequencing to Determine Fitness Effects of Mutations
8:44
Results: High Throughput Sequencing from Whole-Protein Saturation Mutagenesis
10:16
Conclusion
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