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Contents 1 Protocols 1.1 PCR 1.2 Gel Verification 1.3 CRISPR Repeat-Spacer Array Assembly 1.4 Assembling Parts from Oligonucleotides 1.5 (Multi) Site Directed Mutagenesis 1.6 Oligonucleotide Phosphorylation 1.7 Electrocompetent Cell Production 1.8 Gas chromatography-mass spectrometry for biosynthetic product detection 1.9 Make Combinatorial Part Libraries

Protocols

PCR

Instruction for colony PCR and regular PCR

Gel Verification

Make a gel and conduct gel elecrophoresis to varify the presence of inserts

CRISPR Repeat-Spacer Array Assembly

Assemble combinatorial libraries of repeat-spacer-repeat arrays using oligonucleotides

Assembling Parts from Oligonucleotides

Assemble short parts from annealed oligonucleotides

(Multi) Site Directed Mutagenesis

Site directed mutagenesis. Protocol is directly amenable simultaneously making distant mutations.

Oligonucleotide Phosphorylation

Phosphorylate oligonucleotides prior to ligation

Electrocompetent Cell Production

Gas chromatography-mass spectrometry for biosynthetic product detection

GC-MS Protocol for assaying biosynthetic product production.

Make Combinatorial Part Libraries

Make combinatorial part libraries from compatible parts