Template:Team:KULeuven/20 August 2009/VanillinProduction

1)
 * EF colonies present! So, we took 1ml off for further growth and miniprepped the remaining 4ml in triple
 * Nanodrop concentrations:


 * We performed a digest with EcoRI and SpeI to cut out the insert and check it's length. Enzymes were added separately, with 1h in between. Total volume was 30µl.
 * Calculations restriction


 * In the afternoon, the remaining 1ml was re-plated and put on 37°C

2)
 * Test 1 : Results from yesterday's restriction test were inconclusive, because the DNA ladder didn't run properly so the length of the fragments couldn't be read. Therefore, we did it again...
 * Restriction=OK! Test 1 completed.

3)
 * Test 2 : cutting with 2 enzymes subsequently (1h incubation in between) in a volume of 30µl. Using a larger volume should dilute the glycerol, which inhibits cutting. After that, the products are incubated overnight to allow full restriction. sam8 and fcs are cut with EcoRI and SpeI; sam5 and ech are cut with EcoRI and XbaI


 * Calculations restriction