Revision as of 20:08, 25 October 2010 by Theotime (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)

Stéphane
Gel electrophoresis (agarose 1,0% w/v, EtB) Verification of the DNA quantity at each step from the last batch (minipreps, restriction and ligation of 07/15).

• 6 µL DNA ladder (C#83 µg/mL)
• 5 µL pLux (R0062) digest with SpeI and PstI + 1 µL loading buffer
• nothing
• 5 µL pLux (R0062) after gel extraction + 1 µL loading buffer
• 5 µL RBS/GFP/term (E0240) digest with XbaI and PstI + 1 µL loading buffer
• nothing
• 5 µL RBS/GFP/term (E0240) after gel extraction + 1 µL loading buffer
• 5 µL pLux - RBS/GFP/term ligation + 1 µL loading buffer

<a href="/wiki/Image:Verificationgel.jpg" class="image" title="Image:verificationgel.jpg"><img alt="Image:verificationgel.jpg" src="/images/thumb/d/d6/Verificationgel.jpg/334px-Verificationgel.jpg.png" width="334" height="371" border="0" /></a>

Overnight culture
In 10ml LB and 10 µL corresponding antibiotic

• attC - Kan
• term-attC-mRFP - Kan
• term (B0015) - Amp
• RBS/GFP/term (E0240) - Amp
• RBS/LuxR (J37033) - Amp
• pLux (R0062) - Amp
• RBS/LuxI (K081008) - Amp
• Pstrong (J23119) - Amp
• Pmedium (J23110) - Amp
• kanR (P1003) - Amp