Uppsala-Sweden/4 August 2009

From 2009.igem.org

Ethanol Evolution

1,15ml of 95 EtOH were added to a 26ml flask from the recovery protocol of the ethanol evolution project. Thus resulting in approx. 4% EtOH concentration (0,03981% in theory) but I expect that some water evaporated during the culturing thus the former volume (26ml) was decreased. Flask were incubated in 30°C growth room on 120rpm shaker.

Butanol Evolution

26ml flasks from the Butanol recovery were supplied with isobutanol regarding their former concentration. (0,25% | 0,25% | 0,5% | control w/o). Flask were incubated in 30°C growth room on 120rpm shaker.

New Pir A Primers

New Primers for the Pir A gene was ordered Media:PirA primers v_2.xls

PCR (Kivd, ADH2 and PpetE) and Gel run

This is continued since yesterday.

We ran the PCR products on a gel with the following setup and result.

Setup: Media:gel_run_090804_adh2_kivd_ppete.xls

Result:

As clearly seen the PCR failed for all but the PpetE. For kivd the straightforward explanation is absence of the correct strain of lactococcus, although reasons such as poor preparation cannot be ruled out due to the lack of a control (bad planning). In the case of ADH2 a likely cause of failure is the toughness of the yeast cell, preventing exposure of the genome without special lysis being performed.

Thus we will try to find other sources for the correct Lactococcus and repeat the adh2 after lysis of the yeast cells. --Flormane 12:32, 4 August 2009 (UTC)

PCR for transformation

genes: PDC,ADH2 from z.mobilis, PirB, ADH2 from Bakers/Wine yeast and PpetE)

A PCR run was started to make products for the transformation.

This was the setup of annealing Tm and primer design: Media:primers_090804_adh2z_adh2y_pirb_pdc_ppete.xls --Flormane 16:47, 4 August 2009 (UTC)