Difference between revisions of "Team:Bio Without Borders/Notebook"
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<dd>•Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.</dd> | <dd>•Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.</dd> | ||
<dd>•Made linearized backbone from pSB1C3 and pSB1A3 with Phusion PCR</dd> | <dd>•Made linearized backbone from pSB1C3 and pSB1A3 with Phusion PCR</dd> | ||
| − | <dd>•Ran a gel for pSB1A3 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 digest for pSB1C3 linearized backbone and circular (made into linear) pSB1A3 and pSB1C3</dd> | <dd>•Purified Phusion digest for pSB1C3 linearized backbone and circular (made into linear) pSB1A3 and pSB1C3</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> | ||
Revision as of 16:41, 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 PCR using Q5 mastermix.
- •Amplification of pSB1C3 plasmid backbone.
- •Plasmid prep of pSB1A3
- •Digest of linearized pSB1C3
- •Set up phusion PCR and gel electrophoresis of pSB1C3 linearized backbone, using primers VR and VF2.
- •Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.
- •Made 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 Phusion digest for pSB1C3 linearized backbone and circular (made into linear) pSB1A3 and pSB1C3
- Week 5 (July 2-9)
- •Went to Jones Beach to examine horseshoe crabs' habitat and found a carcass