Help:3A Assembly Kit

Revision as of 15:21, 27 April 2012 by Vinoo (Talk | contribs)

3A-Assembly-Kit.jpg

You've just received the 3A Assembly Kit! We hope your team will enjoy using the kit and in the process learn about synthetic biology based on standard parts and 3A assembly. Before you start please make sure to...

  • check the contents of the box with the inventory, to make sure you have everything
  • inspect the materials to make sure nothing has been damaged during shipping
  • refrigerate the agar plates and any materials labelled as requiring storage at 4C.
  • order the NEB reagent kit. You'll need to use the NEB enzymes and reagents for 3A assembly. See the [http://2012hs.igem.org/Sponsors High School Division sponsor page] for more information.


Introduction

The 3A Assembly Kit and protocol will take you through the process of 3A Assembly, and by the end you will have assembled your own composite part in the lab. The kit includes two parts: Part A (BBa_J04500) and Part B (BBa_J04650), which when assembled together will form a composite part device that will make your cells turn red.

The protocol will take you through the following steps:

  1. Growing up the E. coli that contains your parts
  2. Miniprepping the cell cultures to extract the DNA for your parts
  3. Cutting the DNA for your parts and the pSB1C3 linearized plasmid backbone with a restriction digest
  4. Ligating your two parts together into the pSB1C3 linearized backbone
  5. Transforming the ligation to get your newly assembled composite part. If you're successful, your cells will be red.

Note: The 3A Assembly Kit also includes prepared plasmid DNA for your Parts A and B, so you can skip directly to step 3 if you'd like.