Template:Team:KULeuven/20 August 2009/VanillinProduction

From 2009.igem.org

(Difference between revisions)
 
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'''2)'''
'''2)'''
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* [[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...
+
* <u>Test 1</u>: 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.
* Restriction=OK! Test 1 completed.
'''3)'''
'''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
+
* <u>Test 2</u>: 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
* Calculations restriction

Latest revision as of 11:37, 14 September 2009

1)

  • EF colonies present! So, we took 1ml off for further growth and miniprepped the remaining 4ml in triple
  • Nanodrop concentrations:
Part concentration (ng/μl) 260/280 λ
EF A 74,9 1,89
EF B 63,1 1,87
EF C 53,9 1,90
  • 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
Part µl DNA µl MQ
EF C 9,3 20,7
  • 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
Part µl DNA µl MQ
Sam8 B 7,9 22,1
Sam5 B 4,0 26,0
Ech B 5,2 24,8
Fcs B 4,4 25,6
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