Team:NEAU-China/Interlab
1.Who did the actual work to acquire these measurements?
Kang Yang, Cheng Li and Longzhi Cao.
2.What other people should be credited for these measurements? (i.e., who would be an author on any resulting publication. For example, your faculty advisor may have helped design the protocols that you ran.)
Huimin Yue .
3.On what dates were the protocols run and the measurements taken? (this will often be a range of dates; make sure you say which data was taken at what times.)
We ran the protocol on2015.7.16. The data made by flow cytometer was got on 2015.7.21.
4.Do all persons involved consent to the inclusion of this data in publications derived from the iGEM interlab study?
Yes.
1.What protocol did you use to prepare samples for measurement?
We used the Biobrick protocol to assembly and the strain was DH5 Alpha.
2.What sort of instrument did you use to acquire measurements?
o What is the model and manufacturer?
The flow cytometer is Airall.
o How is it configured for your measurements? (e.g., light filters, illumination, amplification)
The the flow cytometer's exciting light was set as 488nm.
3.What protocol did you use to take measurements?
The transformed E coli. Was shaken cultured about 17h and we obtain fluorescence data by the flow cytometer.
4.What method is used to determine whether to include or exclude each sample from the data set?
We use the flow cytometer to exclude some low-activity cell.
5.What exactly were the controls that you used?
DH5 Alpha without transforming which had been shaken cultured for 17h.
6.What quantities were measured? (e.g., red fluorescence, green fluorescence, optical density)
Green fluorescence. For the three reporters are all GFP.
7.How much time did it take to acquire each set of measurements?
It took about 10 minutes to obtain fluorescence data by the flow cytometer.
8.How much does it cost to acquire a set of measurements?
It cost about 500RMB to use the flow cytometer.
9.What are the practical limits on the number or rate of measurements taken with this instrument and protocol?
There is an upper limit to the fluorescence that the machine can record.
<p> <p>1.For each type of quantity measured (e.g., fluorescence, optical density), report on the following: <p>
2.Units:
o What are the units of the measurement? (e.g., meters, molecules)
The number of light spot.
o What is the equivalent unit expressed as a combination of the seven SI base units? (http://en.wikipedia.org/wiki/SI_base_unit)
We just calculated the number, it does not need the SI unit.
3.Precision:
o What is the range of possible measured values for this quantity, using your instrument as configured for these measurements? (e.g., a meter stick measures in the range of 0 to 1 meter)
The spot number is always rational number while the RGB G value range is 0-255.
o What are the significant figures for these measurement? (e.g., on a meter stick, it is common to measure to the nearest millimeter).
Both the number and G value has the precision of 1.
o Is the precision the same across the entire range? If not, how does it differ?
Yes.
o How did you determine these answers?
Our method determined that we couldn't get a discrete result like 0.002. The method is counting instead of detect.
4.Accuracy:
o When was the instrument last calibrated?
2014.8.11.
o How was the instrument calibrated?
Calibrating the light source by raster correction
<p>
<p> <p> <p>1.For each sample, report:
- o the identity of the sample
- o each quantity directly measured
They are saved as picture in the .rar file.
- o each quantity derived from measurements (e.g., fluorescence/OD)
" alt="" width="654" height="272" /><p>2.For each group of replicates, report:
- o the identity of samples in the set
- o which, if any, of the samples are excluded and why
No excluded sample.
- o the mean and standard deviation for each quantity measured or derived