Team:UCLA/Notebook/Honeybee Silk
Honeybee Silk Notebook
April 26 - May 2
4/26 PCR off honey bee gene block
- See Benchling for the g block sequence and primer reference code https://benchling.com/s/p7bClpzQ/edit
- The goal of this is to clone 2 different constructs and prepare them as biobricks, one that is just the silk gene, and the other that as the promoter, rbs, etc.. required for protein expression.
- Using primers p3, p7, and p8. (See Benchling link)
- PCR Reaction 1 (q5 polymerase kit): primers honeybee p#7 and honeybee p#8 with gBlock, and honeybee p#3 and honeybee p#8 amplify just the honeybee coding region and the prom + coding region respectively
Component Volume 5X Q5 Reaction Buffer 5 10 mM dNTPs 0.5 10 uM Forward (primer 3/7) 1.25 10 uM Reverse (primer 8) 1.25 Template (diluted to 1ng/uL) 0.5 Q5 High Fidelity DNA Polymerase 0.25 Nuclease Free Water 16.25 - Had a slice of pizza
May 3 - May 8
5/4 Transformation of Silk and Prom + Silk
- Using Sri’s chemically competent cells
- Using ligation product from 4/30 (not purifying the ligation product as per Eric’s recommendation)
- Restriction digest
- Had a slice of pizza
Day 2
- PCR
- Ran gel
- Restriction digest
- item 1
- item 2
- item 3
- item 4
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Day 3
- PCR
- Ran gel
- Restriction digest
- Had a slice of pizza