A Tour of Addgene's Most Popular Pooled Libraries

By Emily P. Bentley

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Stereotypical UV absorbance spectra exhibited by DNA and other substances commonly used in DNA purification procedures.
Comparison of shRNA and miRNA processing pathways.
Resistance plasmids are depicted with a shield for defense. Col plasmids are depicted with a sword for offense. Virulence plasmids are depicted with a flask of toxins for infection. Degradative plasmids are depicted with a trash can on a dinner plate for hardiness. Fertility plasmids are depicted with a handshake for sharing. Cryptic plasmids are depicted with a question mark for mystery.
Schematic of multicomponent assembly with SLIC.
Schematic of Fragmid assembly process from day one to day four and resulting vector architecture.  On day one fragment plasmids plus a destination vector are assembled using Golden Gate (BbsI). On day two the assembled vector containing a Guide (2xBsmBI), Promoter, N-terminus, Cas protein, C’terminus, and 2A-Selection undergoes an exonuclease V cleanup followed by transformation and plating. On day three two colonies per construct are picked, miniprepped and restriction digested for gel validation/whole plasmid sequencing. Below the assembled vector are examples of six vector architectures including pRDA_512 lentivirus, pRDA_722 lentivirus, pRDA_789 CROPseq lentivirus, pRDA_889 AAV, pRDA_575 Piggyback, and pRDA_791 empty plasmid.

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