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

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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.
 
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.
  
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Methods: We first spoke with Fasih to learn how to order gBlocks and to confirm our primer design methodology. This is what we learned:
+
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
 
1. select gBlocks gene fragments on the IDT (integrated DNA technologies) website
 +
 
2. name the order
 
2. name the order
  
 
3. copy the gene from benchling into the sequence window
 
3. copy the gene from benchling into the sequence window
 +
 
4. test the complexity to make sure its good to go
 
4. test the complexity to make sure its good to go
 +
 
5. say no to all the questions asked by IDT
 
5. say no to all the questions asked by IDT
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Notes:
+
 
 +
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.

Revision as of 00:26, 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.