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Contents 1 Protocols 1.1 PCR 1.2 Gel Verification 1.3 BioBrick Restriction Digest 1.4 BioBrick Ligation 1.5 Making Plates 1.6 Liquid media 1.7 Competent cells 1.8 Transformation 1.9 CRISPR Repeat-Spacer Array Assembly 1.10 Assembling Parts from Oligonucleotides 1.11 (Multi) Site Directed Mutagenesis 1.12 Oligonucleotide Phosphorylation 1.13 Electrocompetent Cell Production 1.14 Gas chromatography-mass spectrometry for biosynthetic product detection 1.15 Make Combinatorial Part Libraries

Protocols

PCR

Instruction for colony PCR and regular PCR

Gel Verification

Make a gel and conduct gel elecrophoresis

BioBrick Restriction Digest

Restriction digest with BioBrick plasmid an inserts

BioBrick Ligation

Ligate Biobrick insert to vector

Making Plates

Make LB plates with antibiotics

Liquid media

Make liquid LB media

Competent cells

Preparation of chemically competent cells

Transformation

Transformation of plasmi DNA into chemically competent cells

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