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

m
m
 
(29 intermediate revisions by 3 users not shown)
Line 1: Line 1:
−
<span style="font-size: 14pt; color: red; font-weight: bold;"><center> We're currently hard at work on the 3A Assembly Kit manual. We've added preliminary protocols for all of the necessary steps so you can read through them for now!</center></span>
+
{{HelpPage/MainLinks}}
 +
{{Template:HelpPage/Discontinued|reason=We are no longer providing the 3A Assembly Kit (as of the 2015 season).}}
  
−
[[Image:3A-Assembly-Kit.jpg|frame|center]]
+
{{Help/3AAK_Style}}
−
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...
+
<html>
−
*check the contents of the box with the [[Help:3A_Assembly_Kit/Inventory|inventory]], to make sure you have everything
+
<table width="100%">
−
*inspect the materials to make sure nothing has been damaged during shipping
+
<tr>
−
*refrigerate the agar plates and any materials labelled as requiring storage at 4C.
+
<td class="headerstep">
−
*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.
+
The 3A Assembly Kit
−
 
+
</td>
−
----
+
<td rowspan=2>
 +
<img src="https://static.igem.org/mediawiki/2013hs/9/91/3A_Assembly_and_Transformation_Efficiency_kits.jpg" style="height: 300px; border: 1px solid #000;">
 +
</td>
 +
</tr>
 +
<tr>
 +
<td style="vertical-align: top; padding: 10px;">
 +
<b>Hi High Schoolers,</b>
 +
<p>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...</p>
 +
<ul>
 +
<li>check the contents of the box with the <a href="https://parts.igem.org/Help:3A_Assembly_Kit/Inventory">inventory</a>, to make sure you have everything</li>
 +
<li>inspect the materials to make sure nothing has been damaged during shipping</li>
 +
<li>refrigerate the agar plates and any materials labeled as requiring storage at 4C. </li>
 +
<li>order an NEB enzyme kit. Click <a href="https://www.neb.com/products/E0546-BioBrick-Assembly-Kit">here</a> for details.
 +
</ul>
 +
<br>
 +
You can download a <a href="https://static.igem.org/mediawiki/parts/e/e9/3A-Assembly-Kit-HiRes.png">hi-res png</a> or <a href="https://static.igem.org/mediawiki/parts/0/0e/3A-Assembly-Kit.pdf">pdf</a> of the images in the protocol.
 +
</td>
  
 +
</tr>
 +
</table>
 +
</html>
  
 
===Introduction===
 
===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''' (<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 red!  
+
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===
 +
 
 +
<html>
 +
<table style="margin-left: auto; margin-right: auto;">
 +
<tr>
 +
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Growing">Step #1: Growing</a></td>
 +
</tr>
 +
<tr>
 +
<td class="introstep">
 +
<p><strong>Grow up the E. coli that contains your parts!</strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Growing">Click here for the Growing Protocol</a></p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/8/8a/3AAK-Diagram-Mini-Grow.png" class="picstep"></td>
 +
</tr>
 +
 
 +
<tr>
 +
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Miniprep">Step #2: Miniprep</a></td>
 +
</tr>
 +
<tr>
 +
<td class="introstep">
 +
<p><strong>Miniprep cell cultures to extract the DNA for your parts.</strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Miniprep">Click here for the Miniprep Protocol</a></p>
 +
</p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/3/3b/3AAK-Diagram-Mini-MP.png" class="picstep"></td>
 +
</tr>
 +
 
 +
<tr>
 +
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Restriction_Digest">Step #3: Restriction Digest</a></td>
 +
</tr>
 +
<tr>
 +
<td class="introstep">
 +
<p><strong>Cut the DNA for your parts and the pSB1C3 linearized plasmid backbone in a restriction digest</strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Restriction_Digest">Click here for the Restriction Digest Protocol</a></p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/b/b5/3AAK-Diagram-Mini-RD.png" class="picstep"></td>
 +
</tr>
 +
 
 +
<tr>
 +
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Ligation">Step #4: Ligation</a></td>
 +
</tr>
 +
<tr>
 +
<td class="introstep">
 +
<p><strong>Ligate your two parts together into the pSB1C3 linearized backbone</strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Ligation">Click here for the Ligation Protocol</a></p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/7/75/3AAK-Diagram-Mini-Lig.png" class="picstep"></td>
 +
</tr>
 +
 
 +
<tr>
 +
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Transformation">Step #5: Transformation</a></td>
 +
</tr>
 +
<tr>
 +
<td class="introstep">
 +
<p><strong>Transform the ligation into E. coli to get your newly assembled composite part.</strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Transformation">Click here for the Transformation Protocol</a></p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/4/45/3AAK-Diagram-Mini-Trf.png" class="picstep"></td>
 +
</tr>
  
−
The protocol will take you through the following steps:
+
<tr>
−
#'''[[Help:3A_Assembly_Kit/Growing|Growing]]''' up the E. coli that contains your parts
+
<td class="headerstep" colspan=2><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Results">Post your Results!</a></td>
−
#'''Miniprepping''' the cell cultures to extract the DNA for your parts
+
</tr>
−
#Cutting the DNA for your parts and the pSB1C3 linearized plasmid backbone in a '''restriction digest'''
+
<tr>
−
#'''Ligating''' your two parts together into the pSB1C3 linearized backbone
+
<td class="introstep">
−
#'''Transforming''' the ligation to get your newly assembled composite part. At the end you should see red colonies!
+
<p><strong>Post your results! Hopefully you've learned quite a bit while using the 3A Assembly Kit. </strong></p>
 +
<p><a href="https://parts.igem.org/Help:3A_Assembly_Kit/Results">Click here to post your Results!</a></p>
 +
</td>
 +
<td><img src="https://static.igem.org/mediawiki/parts/8/85/3AAK-Diagram-Mini-Res.png" class="picstep"></td>
 +
</tr>
 +
</table>
 +
</html>
  
−
'''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.
+
[[Category:Protocols]]

Latest revision as of 16:51, 1 June 2017

This page has been discontinued. We apologize for any inconvenience. For more information please contact iGEM HQ

Reason: We are no longer providing the 3A Assembly Kit (as of the 2015 season).


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!