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Contents 1 Protocols 1.1 CRISPR Repeat-Spacer Array Assembly 1.2 Assembling Parts from Oligonucleotides 1.3 (Multi) Site Directed Mutagenesis 1.4 Oligonucleotide Phosphorylation 1.5 Electrocompetent Cell Production

Protocols

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

Make electrocompetent cells.