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Revision as of 16:39, 22 July 2014

https://parts.igem.org/Help:

The 3A Assembly Kit
Hi High Schoolers,

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 labeled as requiring storage at 4C.
  • order an NEB enzyme kit. Click here for details.

You can download a hi-res png or pdf of the images in the protocol.

Introduction

The 3A Assembly Kit and following protocols 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 RFP (red fluorescent protein) generator. Your cells will turn red!

Note: The 3A Assembly Kit also includes prepared plasmid DNA for Part A and Part B, so you can skip directly to Step #3: Restriction Digest if you'd like. But don't forget to take a look at Step #1: Grow, as you'll need to follow the instructions for growing up cells for Step #5: Transformation.

Protocols

Step #1: Growing

Grow up the E. coli that contains your parts!

Click here for the Growing Protocol
Step #2: Miniprep

Miniprep cell cultures to extract the DNA for your parts.

Click here for the Miniprep Protocol
Step #3: Restriction Digest

Cut the DNA for your parts and the pSB1C3 linearized plasmid backbone in a restriction digest

Click here for the Restriction Digest Protocol
Step #4: Ligation

Ligate your two parts together into the pSB1C3 linearized backbone

Click here for the Ligation Protocol
Step #5: Transformation

Transform the ligation into E. coli to get your newly assembled composite part.

Click here for the Transformation Protocol
Post your Results!

Post your results! Hopefully you've learned quite a bit while using the 3A Assembly Kit.

Click here to post your Results!