Plasmids (2)

Recent Posts

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.
A cartoon schematic of prokaryotic chromosomal replication. The parent cell DNA is shown as a circular chromosome with a small region highlighted as the origin of replication, or ori.   During replication, the DNA helix has separated at the ori, creating a “bubble” of two single strands of DNA. The point of separation of the helix into these single strands is the replication fork. Two replication forks form on either end of the ori. At each replication fork, a helicase processively separates the DNA strands, and a polymerase synthesizes a new DNA strand paired to each single parent strand.  After termination, which is not shown in detail, the process results in two identical daughter cell DNA chromosomes.
Cartoon depicting factors to consider for a pooled library when creating an amplification protocol; details are in text of link.
A chart with MoClo kit recommendations for different modal organisms and applications. 
Graphic representation of the FITS screening process
Graphic showing a narrow range host plasmid with one ORI, and a broad range host plasmid with three ORIs with variable inserts.
Schematic showing the three assembly levels of MoClo. Level 1 has 7 individual parts on a plasmid; Level 1 has seven parts all together on on plasmid; Level 2+ has three sets of five parts each on a plasmid, with two additional parts on the plasmid but separated from the sets.

Sharing science just got easier... Subscribe to our blog