Module 2, lab 6 - GAPC gene cloningPlasmid DNA purification ('miniprep')

Today we used the E. coli cultures Salesha and Jess inoculated yesterday to do a small-scale plasmid DNA extraction ('miniprep') using Promega's Wizard® Plus SV Minipreps DNA Purification System. Then restriction enzyme digestions with Bgl II were set to confirm that the insert (GAPC gene) is present in the plasmid (pJet1.2).

(Section 2 did the procedure on Monday [MLK day]. Thanks to Stacy for inoculating the media the day before!)

Here's a recap of what we have done so far in the last couple of exercises:

Lab 5 - Nested PCR of the GAPC gene from Arabidopsis

Lab 6 - Cloning he GAPC gene from Arabidopsis

• Ligation of PCR amplified GAPC gene onto the pJet1.2 plasmid
• Genetic transformation of E. coli with the pJet1.2 plasmid
• Cloning of genetically transformed E. coli
• Minipreps (purification of pJet1.2 plasmid)

Next, running an agarose gel to confirm the results of the R.E.D. (lab 6) and setting DNA sequencing reactions (lab 7)

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