diff --git a/wiki/pages/protocols.html b/wiki/pages/protocols.html index 9c158fa8a0134f0cc002b38e51c6cfe53c51ea74..f42518f95b19b3138ae222e9f00c8bc74994597c 100644 --- a/wiki/pages/protocols.html +++ b/wiki/pages/protocols.html @@ -479,7 +479,7 @@ with 10µl liquid culture onto YPD(+2%Glucose) solid<br> -Incubate at 30°C in stationary incubator for 2-3 days in absence of light<br><span class="contentprotocolsubhead">3. Streaking×2</span><br>-Inoculate single colonies onto a SC-Ura plate first (via streaking), without changing pipette tip, streak again on a YPD(+2%Glucose) plate<br> -Incubate at 30°C in stationary incubator for 2-3 days<br><span class="contentprotocolsubhead">4. Genome Sequencing</span><br>-Yeast that can not grow on SC-Ura have their plasmids completely eliminated, send the according yeast colonies on YPD plates for sequencing<br> - <br><span class="contentprotocolsubhead">Saccharomyces Culturing</span><br> <span class="contentprotocolsubhead">5. Inoculation</span><br>-Inoculate a colony with correct sequencing results into 5ml liquid YPD(+2%Glucose)<br> -Incubate at 28°C to 30°C and 280RPM overnight<br><span class="contentprotocolsubhead">6. Pre-Culture</span><br>-Add 50ml liquid YPD(+2%Glucose) to sterilized 100ml conical flask<br> -Inoculate 500µl liquid culture into the + <br><span class="contentprotocolsubhead"><i>Saccharomyces</i> Culturing</span><br> <span class="contentprotocolsubhead">5. Inoculation</span><br>-Inoculate a colony with correct sequencing results into 5ml liquid YPD(+2%Glucose)<br> -Incubate at 28°C to 30°C and 280RPM overnight<br><span class="contentprotocolsubhead">6. Pre-Culture</span><br>-Add 50ml liquid YPD(+2%Glucose) to sterilized 100ml conical flask<br> -Inoculate 500µl liquid culture into the conical flask, incubate at 30°C and 220 RPM at 1.5 to 2 hours so that OD600 reaches 0.4 to 0.6<br><span class="contentprotocolsubhead">7. Culture</span><br>Centrifuge pMVA+SCIE8 culture at 2000RPM for 5 minutes, carefully transfer the dodecane layer into yeast liquid culture, culture at 30°C and 220RPM overnight<br><span class="contentprotocolsubhead">8. GC-MS Analysis</span><br>-Transfer overnight liquid culture into a centrifuge tube<br> -Centrifuge at 12000RPM for 5 minutes<br> -Transfer and filter the layer of dodecane into GC-MS sample tube<br> -Mailing for analysis<br> @@ -489,7 +489,7 @@ </div> <div class="contentBx"> - <div class="label">Co-culture of yeast and E.coli</div> + <div class="label">Co-culture of <i>S. cerevisiae</i> with <i>E. coli</i></div> <div class="contentprotocol"> <p> <strong>Note: </strong>All operations must be conducted in a clean bench! Please ensure sterile techniques.<br> @@ -531,7 +531,7 @@ </ul> </p> <p>YPD + 2% Glucose: 47.5 ml YPD + 2.5 ml 40% glucose<br> - <br> <span class="contentprotocolsubhead">Day 5:</span><br> <span style="color: #000;font-size: 20px;font-weight: 600;">Co-culture of E. coli and Yeast</span><br> 1. Dilute the yeast culture 10 times, measure the yeast culture + <br> <span class="contentprotocolsubhead">Day 5:</span><br> <span style="color: #000;font-size: 20px;font-weight: 600;">Co-culture of <i>E. coli</i> and <i>S. cerevisiae</i></span><br> 1. Dilute the yeast culture 10 times, measure the yeast culture OD600, and calculate the required amount of yeast culture. <ul> <li> Control: 2000 µl YPD; Sample: 1800 µl YPD + 200 µl yeast culture</li> @@ -539,14 +539,14 @@ </ul> </p> - <p>2. Transfer the calculated amount of yeast culture to a centrifuge tube, centrifuge at 6000 g for 10 minutes, and carefully discard the supernatant.<br>3. Resuspend the yeast pellet in 48h cultured E. coli liquid, gently mix by - pipetting, and incubate the mixture of E. coli and yeast at 28°C, 200 rpm for 48 hours. + <p>2. Transfer the calculated amount of yeast culture to a centrifuge tube, centrifuge at 6000 g for 10 minutes, and carefully discard the supernatant.<br>3. Resuspend the yeast pellet in 48h cultured <i>E. coli</i> liquid, gently mix by + pipetting, and incubate the mixture of <i>E. coli</i> and yeast at 28°C, 200 rpm for 48 hours. </p> <br> <hr style="width: 100%;"> <br> <p> - <span class="contentprotocolsubhead">Day 7:</span><br> 1. Transfer the mixed culture of E. coli and yeast to a centrifuge tube, centrifuge at 12000 rpm for 5 minutes to collect the culture.<br> 2. Add 5% dodecane to the collected + <span class="contentprotocolsubhead">Day 7:</span><br> 1. Transfer the mixed culture of <i>E. coli</i> and yeast to a centrifuge tube, centrifuge at 12000 rpm for 5 minutes to collect the culture.<br> 2. Add 5% dodecane to the collected culture, perform extraction, let it stand to allow separation, and then take the dodecane layer to filter into a GC-MS sample vial. Send for GC-MS analysis.<br> <br><span style="color: #000;font-size: 20px;font-weight: 600;">Reagent Preparation:</span>