Line 31: | Line 31: | ||
<dd>•Created a 50mg/mL kanamycin solution and then alloquoted it to about 80 1.5mL tubes. </dd> | <dd>•Created a 50mg/mL kanamycin solution and then alloquoted it to about 80 1.5mL tubes. </dd> | ||
<dd>•Inoculated LB with ampicillin colonies (with insert).</dd> | <dd>•Inoculated LB with ampicillin colonies (with insert).</dd> | ||
− | <dd>•Streaked ampicillin ( | + | <dd>•Streaked ampicillin (1A3) colony onto new plates.</dd> |
− | <dd>•Conducted cPCR for | + | <dd>•Conducted cPCR for 1A3 colonies with insert.</dd> |
<dt><b><font color="#009999">Week 4 (June 25-29)</font></b></dt> | <dt><b><font color="#009999">Week 4 (June 25-29)</font></b></dt> | ||
<dd>•Conducted cPCR for PSB1K3.</dd> | <dd>•Conducted cPCR for PSB1K3.</dd> | ||
Line 41: | Line 41: | ||
<dd>•Amplification of PSB1C3 plasmid backbone.</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>•TMR!!! Amplification of psb1c3 and psb1a3, ran it through PCR cleanup and digested with | + | <dd>•TMR!!! Amplification of psb1c3 and psb1a3, ran it through PCR cleanup and digested with EcoR1 and Pst1.</dd> |
Revision as of 21:49, 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.