Team:UC Berkeley/GatewayPlasmid


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Gateway Plasmid Schemes

Scheme 1: Gateway Machinery in Assembly Vector

Assembly Vectors:Two different variations of the pK112128 assembly vector were created: one with {!} and one with {!}{rbs.ihfA!}{rbs.ihfB!} between the ccdB site and the attR2 site. These plasmids are named pK112245 and pK112246 respectively (see images below).


Entry Vector:The entry vector used in both cases was pBca1256 (see image below).
PBca 1256.png

Scheme 2: Gateway Machinery in Entry Vector

Assembly Vector:The assembly vector used for each of the following entry vector variations is pK112128 (see image below.

Entry Vectors: Two different variations of the pBca1256 entry vector were created: one with {!} and one with {!}{rbs.ihfA!}{rbs.ihfB!} just before the attL1 site. These plasmids are named pK112247 and pK112248 respectively (see images below).


General Procedures

  1. Transform 1ul of mini-prepped assembly vector DNA plasmid into TG1 cells
  2. Rescue for 45 minutes at 30 deg C
  3. Plate