"
Uppsala-Sweden/16 July 2009
From 2009.igem.org
Karl.brune (Talk | contribs) |
|||
| (One intermediate revision not shown) | |||
| Line 1: | Line 1: | ||
| - | '''Starting up | + | '''Starting up ethanol and butanol survival experiments''' |
Goal: Measure the ethanol and isobutanol tolerance of Synecocystis. Will be done using triplicates of ethanol in E-flasks and 5x replicates of ethanol and isobutanol on 96-well plates. | Goal: Measure the ethanol and isobutanol tolerance of Synecocystis. Will be done using triplicates of ethanol in E-flasks and 5x replicates of ethanol and isobutanol on 96-well plates. | ||
| Line 24: | Line 24: | ||
/Anders | /Anders | ||
| + | |||
| + | '''Recovery of Z. mobilis ATCC 29191 ''' | ||
| + | The Z. mobilis ATCC 29191 strain from the FZ Jülich, Germany arrived today. (Thanks to Dr. Stephanie Bringer) | ||
| + | |||
| + | Strain specific medium* was prepared and the lyphophilisate was inoculated in 15 ml culture tubes and spread out on 5 plates. The plates and the tubes were then incubated at 30°C. The tubes on a 120rpm shaker. | ||
| + | |||
| + | Medium: | ||
| + | |||
| + | Yeast extract 10g | ||
| + | Glucose 55g | ||
| + | KH2PO4 1g | ||
| + | (NH4)2SO4 1g | ||
| + | MgSO4 7H2O 0,5g | ||
| + | dH2O 1000ml | ||
| + | ->(for the plates)+ Agar 15g | ||
| + | |||
| + | --[[User:Karl.brune|Karl.brune]] 20:05, 19 July 2009 (UTC) | ||
| + | /Karl | ||
Latest revision as of 20:05, 19 July 2009
Starting up ethanol and butanol survival experiments Goal: Measure the ethanol and isobutanol tolerance of Synecocystis. Will be done using triplicates of ethanol in E-flasks and 5x replicates of ethanol and isobutanol on 96-well plates.
Volumes: 25 ml final for E-flask and 200 µl final for plates. Concentrations: 8, 4, 2, 1, 0 vol% ethanol for E-flasks and plate, 2, 1, 0,5 , 0,25 , 0 vol% percent isobutanol on plate.
Growth media: BG-11, nitrate supplemented as dictated by standard protocol. Ethanol used 95 vol% supplemented to BG-11 nutritional value.
11:00 OD measurement of start culture. OD found to be 0,124.. at eight times dilution, within linear range as established 09-07-16 (OD 0,334-0,042)
14:00 Butanol experiment fails as unable to pipette the minute volumes of butanol (<10 µl 25% isubutanol) needed at plate
16:00 Inoculation concluded, cultures placed in growth room at 30 degrees Celsius, E-flasks on 120 rpm shaker table and plate on --- rpm shaker table.
First OD measurement, correct amounts of cull culture verified.
17:00 First measurement included in growth curve, no difference in growth detected. Problems with plate from condensation on lid, tries to solve with pre heated plate (35 degrees C) reader.
Still fogging on inside of plate lid, values uncertain.
/Anders
Recovery of Z. mobilis ATCC 29191 The Z. mobilis ATCC 29191 strain from the FZ Jülich, Germany arrived today. (Thanks to Dr. Stephanie Bringer)
Strain specific medium* was prepared and the lyphophilisate was inoculated in 15 ml culture tubes and spread out on 5 plates. The plates and the tubes were then incubated at 30°C. The tubes on a 120rpm shaker.
Medium:
Yeast extract 10g Glucose 55g KH2PO4 1g (NH4)2SO4 1g MgSO4 7H2O 0,5g dH2O 1000ml ->(for the plates)+ Agar 15g
--Karl.brune 20:05, 19 July 2009 (UTC) /Karl
