Difference between revisions of "Help:3A Assembly Kit"

Revision as of 21:43, 4 January 2013

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 the NEB reagent kit. You'll need to use the NEB enzymes and reagents for 3A assembly. See the High School Division sponsor page for more information.

If you're interested in using the 3A Assembly Kit, and would like to order one, please contact hq (at) igem (dot) org. You can also 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!

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 ===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''' (<partinfo>BBa_J04500</partinfo>) and '''Part B''' (<partinfo>BBa_J04650</partinfo>), which when assembled together will form a RFP (red fluorescent protein) generator. Your cells will turn <span style="color: red; font-weight: bold;">red!</span>
+The 3A Assembly Kit and following protocols will take you through the process of [[Help:Assembly#3A_Assembly | 3A Assembly]], and by the end you will have assembled your own composite part in the lab. The kit includes two parts: '''Part A''' (<partinfo>BBa_J04500</partinfo>) and '''Part B''' (<partinfo>BBa_J04650</partinfo>), which when assembled together will form a RFP (red fluorescent protein) generator. Your cells will turn <span style="color: red; font-weight: bold;">red!</span>
 '''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===