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Main purpose

• ON culture of Colony PCR products of HAO and AMO from 19-07-2013
• USER reaction of AMO and HAO with pZA21 (with native promoter).
• RFP amplification in pZA21 using primers without promoter
• PCR to amplify USER-compatible AMO and HAO fragments

Who was in the lab

Gosia, Kristian, Henrike

Results

Verification gel

1% gel to verify our earlier PCR purifications

wells (in bracets position of the purification in the gene box):

• 1: broad band ladder
• 2: HAO extracted from N. europeae with flanking primers (A4)
• 3: AMO extracted from N. europeae with flanking primers (A6)
• 4: cycAX extracted from N. europeae with flanking primers (A5)
• 5: Nir extracted from P. aeruginosa with flanking primers (A7)
• 6: Nir extracted from P. aeruginosa with flanking primers (A8)
• 7: Nir extracted from P. aeruginosa with flanking primers (B8)
• 8: Nir extracted from P. aeruginosa with flanking primers (B9)
• 9: pZA21 USER fragment, DpnI treated (F1)
• 10: cycAX USER fragment (G1)
• 11: AMO USER fragment (G2)
• 12: HAO USER fragment (G3)
• 13: Nir USER fragment (G4)
• 14: AMO USER fragment (G5)
• 15: 1kb plus ladder

colony PCR gel

1% gel to analyse yesterday's colony PCR

wells:

• 1: 1 kb ladder
• 2: HAO colony 1
• 3: HAO colony 2
• 4: RFP in pZA21 without promoter
• 5: RFP in pZA21 without promoter
• 6: AMO colony 3 (count starts from 3)
• 7: AMO colony 4
• 8: AMO colony 5
• 9: AMO colony 6
• 10: AMO colony 7
• 11: AMO colony 8
• 12: AMO colony 9
• 13: AMO colony 10
• break in the gel
• 14: cycAX colony 1
• 15: cycAX colony 2
• 14: RFP colony 1
• 14: RFP colony 2