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<dt><b><font color="#009999">Week 3 (June 18-22)</font></b></dt> | <dt><b><font color="#009999">Week 3 (June 18-22)</font></b></dt> | ||
<dd>•Made chloramphenicol LB plates</dd> | <dd>•Made chloramphenicol LB plates</dd> | ||
− | <dd>•Restriction enzyme digest of iGem plasmid | + | <dd>•Restriction enzyme digest of iGem plasmid pSB1C3 using EcoR1 and Pst1</dd> |
− | <dd>•Made digest master mix; ran an ezyme digest of iGem plasmids | + | <dd>•Made digest master mix; ran an ezyme digest of iGem plasmids pSB1A3 and pSB1K3 |
Performed ligation of </dd> | Performed ligation of </dd> | ||
− | <dd> | + | <dd>pSB1A3 and pSB1K3; transformed pSB1A3 in competent cells on ampicillin plate</dd> |
<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> | ||
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<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> | ||
− | <dd>•Ran 1% agarose gel of | + | <dd>•Ran 1% agarose gel of pSB1A3 with the insert (colonies that were re-streaked).</dd> |
− | <dd>•Ran 1% agarose gel of | + | <dd>•Ran 1% agarose gel of pSB1K3 with insert.</dd> |
<dd>•Cleaned laboratory material. | <dd>•Cleaned laboratory material. | ||
<dd>•Set up PCR using Q5 mastermix.</dd> | <dd>•Set up PCR using Q5 mastermix.</dd> | ||
− | <dd>•Amplification of | + | <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 | + | <dd>•TMR!!! Amplification of pSB1C3 and pSB1A3, ran it through PCR cleanup and digested with EcoR1 and Pst1.</dd> |
Revision as of 21:50, 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.