Difference between revisions of "Team:UCLA/Notebook/Protein Cages/25 June 2015"

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Remember to resuspend the gene at a concentration of 10 ng/microL.  
 
Remember to resuspend the gene at a concentration of 10 ng/microL.  
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Add a double stop codon.  
 
Add a double stop codon.  
 
Snap gene is a good place to look for plasmid sequences.
 
Snap gene is a good place to look for plasmid sequences.
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Tyler Lee
 
Tyler Lee
 
--[[User:Wtleeiv|Wtleeiv]] 19:27, 25 June 2015 (CDT)
 
--[[User:Wtleeiv|Wtleeiv]] 19:27, 25 June 2015 (CDT)
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Introduction:
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During our downtime with either lab protocols or orders to arrive, we decided to do a formal journal club within the protein cages group. Our first journal club will involve PCR(procedure,theory, application) and site directed mutagenesis as these will be heavily used in the project.
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1.I was preparing for this journal club by finding papers and preparing a powerpoint presentation for this.
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2.I attended the meeting with David and Philip to go over our PCquad sequence and to decide what exactly to order.
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Nithin

Revision as of 01:30, 26 June 2015

iGEM UCLA




Intro: Today we designed the primer sequence for our codon optimized cage sequence from Yates' lab. I initially researched the iGEM specifics for the prefixes and suffix they require, so that we knew how to design primers for the sequence. Additionally, another set of primers with NdeI (prefix) and XhoI (suffix) sites will be designed for protein expression within the pET22b vector, since this plasmid was used by Dr. Yates.

We first spoke with Fasih to learn how to order gBlocks and to confirm our primer design methodology. This is what we learned:

1. select gBlocks gene fragments on the IDT (integrated DNA technologies) website

2. name the order

3. copy the gene from benchling into the sequence window

4. test the complexity to make sure its good to go

5. say no to all the questions asked by IDT

6. Notes: A. for payment, use oilgocard B. send to david attn. iGEM C. 1,000 ng is a good guaranteed yield

Remember to resuspend the gene at a concentration of 10 ng/microL.


Add a double stop codon. Snap gene is a good place to look for plasmid sequences.

Tyler Lee --Wtleeiv 19:27, 25 June 2015 (CDT)


Introduction: During our downtime with either lab protocols or orders to arrive, we decided to do a formal journal club within the protein cages group. Our first journal club will involve PCR(procedure,theory, application) and site directed mutagenesis as these will be heavily used in the project.

1.I was preparing for this journal club by finding papers and preparing a powerpoint presentation for this. 2.I attended the meeting with David and Philip to go over our PCquad sequence and to decide what exactly to order.

Nithin