Difference between revisions of "Part:BBa K1679005"

Revision as of 02:10, 19 September 2015 (view source) QinQikai (Talk | contribs) ← Older edit		Revision as of 02:17, 19 September 2015 (view source) QinQikai (Talk | contribs) (→‎Flow Cytometer Result) Newer edit →
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	===Flow Cytometer Result===		===Flow Cytometer Result===
−	<h2>Flow Cytometer Measurement</h2>
	<p></p>		<p></p>
	<h3>Equipment</h3>		<h3>Equipment</h3>
	<p>Epics-XL<BR>Beckman Coulter</p>		<p>Epics-XL<BR>Beckman Coulter</p>
−	<div style="width: 50%;margin: 0 auto;">
−	<center><img src="https://static.igem.org/mediawiki/2015/7/7c/OUC-China-InterLab_6.jpg" alt="" class="img-responsive"></center>
−	</div>
	<h3>Protocol</h3>		<h3>Protocol</h3>
	<p>		<p>
	Date: 2015 Aug 12<BR>1. Start up the instrument and warm up it until it shows “Awaiting Sample”<BR>2. Calibrate the instrument with check beads<BR>3. Mix the fluorescent beads and negative sample (E coli. without plasmid) and dilute it to 1 mL<BR>4. Measure the mix and regulate voltage until the points appear in the appropriate location and set gates<BR></p>		Date: 2015 Aug 12<BR>1. Start up the instrument and warm up it until it shows “Awaiting Sample”<BR>2. Calibrate the instrument with check beads<BR>3. Mix the fluorescent beads and negative sample (E coli. without plasmid) and dilute it to 1 mL<BR>4. Measure the mix and regulate voltage until the points appear in the appropriate location and set gates<BR></p>
	<div style="width: 60%;margin: 0 auto;">		<div style="width: 60%;margin: 0 auto;">
−	<center><img src="https://static.igem.org/mediawiki/2015/d/df/OUC-China-InterLab_9.png" alt="" class="img-responsive"></center>	+	<center>https://static.igem.org/mediawiki/2015/d/df/OUC-China-InterLab_9.png</center>
	</div>		</div>
	<div style="width: 60%;margin: 0 auto;">		<div style="width: 60%;margin: 0 auto;">
−	<center><img src="https://static.igem.org/mediawiki/2015/2/28/OUC-China-InterLab_10.jpg" alt="" class="img-responsive"></center>	+	<center>https://static.igem.org/mediawiki/2015/2/28/OUC-China-InterLab_10.jpg</center>
	</div>		</div>
	<p>5. Measure the other samples in three biological replicates and three technical replicates<BR>6. Use the Cleanase and ddH2O to clean the instrument as instruction<BR>7. Process the data		<p>5. Measure the other samples in three biological replicates and three technical replicates<BR>6. Use the Cleanase and ddH2O to clean the instrument as instruction<BR>7. Process the data
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	<h3>Dataset</h3>		<h3>Dataset</h3>
	<div style="width: 60%;margin: 0 auto;">		<div style="width: 60%;margin: 0 auto;">
−	<center><img src="https://static.igem.org/mediawiki/2015/f/fb/OUC-China-InterLab_11.png" alt="" class="img-responsive"></center>	+	<center>https://static.igem.org/mediawiki/2015/f/fb/OUC-China-InterLab_11.png</center>
	</div>		</div>

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Revision as of 02:17, 19 September 2015

GFP Reporter

This is a device that contain promoter BBa_J23117, RiboJ and BBa_I13504.

Plate Reader Result

Equipment

Varioskan Flash Multimode Reader
Thermo Scientific
Read Speed: 6.4 well per second

Software

Run Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash
Current Software Version: Skanlt Software 2.4.5 RE for Varioskan Flash

Protocol

Date: 2015 Aug 12
1. Set our instrument to read OD600
2. Setup a 96-well plate with our cultures
3. Take the measurement and record it
4. Calculate the dilution required for each sample (OD600=0.5)
5. Dilute each sample
6. Remeasure our sample on OD600
7. Recalculate our dilution and remeasure until it's within 5% of 0.5.

Unit

Three technical replicates of M9 liquid media was measured, which were called background value.
A series of concentration of sodium fluorescein was measured, which are 0, 10, 20, 40, 80, 160, 320, 640 ng/mL . A calibration curve was defined.

Dataset

All samples were cut the mean of background value, and then compared with the calibration curve to get the final dataset in units of fluorescein.

TR: Technical Replicate
BR: Biological Replicate

Flow Cytometer Result

Equipment

Epics-XL
Beckman Coulter

Protocol

Date: 2015 Aug 12
1. Start up the instrument and warm up it until it shows “Awaiting Sample”
2. Calibrate the instrument with check beads
3. Mix the fluorescent beads and negative sample (E coli. without plasmid) and dilute it to 1 mL
4. Measure the mix and regulate voltage until the points appear in the appropriate location and set gates

5. Measure the other samples in three biological replicates and three technical replicates
6. Use the Cleanase and ddH2O to clean the instrument as instruction
7. Process the data

Unit

The fluorescence of GFPwas compared with the 2.00μm Fluoresbrite Yellow Green (YG) Microspheres.

Dataset

Sequence and Features