Revision as of 21:55, 28 June 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.; • Ran colony PCR and plated bacteria on antimicrobial plate.; • cPCR, chloramphenicol, and culturing protocols, made agarose gel; • gel electrophoresis for cPCR, nucleic acid purification plasmid
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.
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.; •Set up phusion PCR and gel electrophoresis of PSB1A3.; •TMR!!! Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.

@@ Line 37: / Line 37: @@
 <dd>•Ran 1% agarose gel of pSB1A3 with the insert (colonies that were re-streaked).</dd>
 <dd>•Ran 1% agarose gel of pSB1K3 with insert.</dd>
-<dd>•Cleaned laboratory material.
+<dd>•Cleaned laboratory material.</dd>
 <dd>•Set up PCR using Q5 mastermix.</dd>
 <dd>•Amplification of pSB1C3 plasmid backbone.</dd>
-<dd>•Set up phusion PCR and gel electrophoresis of PSB1A3.
+<dd>•Set up phusion PCR and gel electrophoresis of PSB1A3.</dd>
 <dd>•TMR!!! Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.</dd>