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<dd>•Ran 1% agarose gel of pSB1K3 with insert.</dd> | <dd>•Ran 1% agarose gel of pSB1K3 with insert.</dd> | ||
<dd>•Cleaned laboratory material.</dd> | <dd>•Cleaned laboratory material.</dd> | ||
− | <dd>•Set up PCR using Q5 | + | <dd>•Set up pSB1C3 backbone amplification PCR using Q5.</dd> |
− | <dd>•Amplification of pSB1C3 | + | <dd>•Ran gel of pSB1C3 j04450 insert with Q5 PCR to confirm size.</dd> |
+ | <dd>•Amplification of pSB1C3 j04450 insert with Phusion PCR (because Q5 did not work). Confirmed size with gel electrophoresis.</dd> | ||
<dd>•Plasmid prep of pSB1A3</dd> | <dd>•Plasmid prep of pSB1A3</dd> | ||
− | <dd>•Digest of | + | <dd>•Digest of amplified pSB1C3 j04450 insert (Phusion PCR).</dd> |
− | + | <dd>•Amplified linearized backbone from pSB1C3 and pSB1A3 with Phusion PCR</dd> | |
− | + | ||
− | <dd> | + | |
<dd>•Ran a gel for pSB1A3 and pSB1C3 plasmid prep from day before to check band size</dd> | <dd>•Ran a gel for pSB1A3 and pSB1C3 plasmid prep from day before to check band size</dd> | ||
− | <dd>•Purified Phusion | + | <dd>•Purified (accidentally-amplified with Phusion) PSB1C3 j04450 insert, as well as the circular (made into linear) pSB1A3 and pSB1C3 backbones with PCR clean-up</dd> |
<dt><b><font color="#009999">Week 5 (July 2-9)</font></b></dt> | <dt><b><font color="#009999">Week 5 (July 2-9)</font></b></dt> | ||
<dd>•Went to Jones Beach to examine horseshoe crabs' habitat and found a carcass</dd> | <dd>•Went to Jones Beach to examine horseshoe crabs' habitat and found a carcass</dd> | ||
+ | <dd>•Worked on GoFundMe campaign page</dd> | ||
Revision as of 17:18, 5 July 2018
Notebook
- Week 1 (June 4-8)
- •Made competent cells
- Week 2 (June 11-15)
- •Testing competent cells using iGem DNA transformation samples.
- • Tested to see if they grew better on a freezer block versus an ice bath. They grow better in an ice bath.
- • Ran colony PCR and plated bacteria on antimicrobial plate.
- • cPCR, chloramphenicol, and culturing protocols
- • Ran 1% agarose gel for cPCR
- • Plasmid prep of pSB1C3
- Week 3 (June 18-22)
- •Made chloramphenicol LB plates
- •Restriction enzyme digest of iGem plasmid pSB1C3 using EcoR1 and Pst1
- •Made digest master mix; ran an ezyme digest of iGem plasmids pSB1A3 and pSB1K3 Performed ligation of
- pSB1A3 and pSB1K3; transformed pSB1A3 in competent cells on ampicillin plate
- •Created a 50mg/mL kanamycin solution and then alloquoted it to about 80 1.5mL tubes.
- •Inoculated LB with ampicillin colonies (with insert).
- •Streaked ampicillin (1A3) colony onto new plates.
- •Conducted cPCR for 1A3 colonies with insert.
- •Filled out first draft of safety form
- •Went to Brighton Beach to examine horseshoe crabs' habitat and found a part of the carcass
- Week 4 (June 25-29)
- •Conducted cPCR for pSB1K3.
- •Ran 1% agarose gel of pSB1A3 with the insert (colonies that were re-streaked).
- •Ran 1% agarose gel of pSB1K3 with insert.
- •Cleaned laboratory material.
- •Set up pSB1C3 backbone amplification PCR using Q5.
- •Ran gel of pSB1C3 j04450 insert with Q5 PCR to confirm size.
- •Amplification of pSB1C3 j04450 insert with Phusion PCR (because Q5 did not work). Confirmed size with gel electrophoresis.
- •Plasmid prep of pSB1A3
- •Digest of amplified pSB1C3 j04450 insert (Phusion PCR).
- •Amplified linearized backbone from pSB1C3 and pSB1A3 with Phusion PCR
- •Ran a gel for pSB1A3 and pSB1C3 plasmid prep from day before to check band size
- •Purified (accidentally-amplified with Phusion) PSB1C3 j04450 insert, as well as the circular (made into linear) pSB1A3 and pSB1C3 backbones with PCR clean-up
- Week 5 (July 2-9)
- •Went to Jones Beach to examine horseshoe crabs' habitat and found a carcass
- •Worked on GoFundMe campaign page