Difference between revisions of "Part:BBa K1679001"

Latest revision as of 01:27, 19 September 2015

GFP Reporter

This is a device containing a promoter (BBa_J23101),a RBS (BBa_B0034), a GFP coding sequence (BBa_E0040) and two terminators (BBa_B0010, BBa_B0012) on the pSB1C3, which can be easily transformed into E.coli and produce GFP (mut3b). This is a GFP expression device, which can express GFP (mut3b) without any induction.

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

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 705

Functional Parameters

chassis	E. coli K-12 DH5-alpha
emission	511nm
excitation	501nm
function	-NA-

@@ Line 18: / Line 18: @@
                  <p>
                      Date: 2015 Aug 12<BR>1. Set our instrument to read OD600<BR>2. Setup a 96-well plate with our cultures<BR>3. Take the measurement and record it<BR>4. Calculate the dilution required for each sample (OD600=0.5)<BR>5. Dilute each sample<BR>6. Remeasure our sample on OD600<BR>7. Recalculate our dilution and remeasure until it's within 5% of 0.5.
-                </p>
-                <h3>Layout</h3>
-                <div style="width: 70%;margin: 0 auto;">
-                    https://static.igem.org/mediawiki/2015/7/7f/OUC-China-InterLab_1.png
-                </div>
-                <p>
-: J23101+I13504; 106: J23106+I13504; 117: J23117+I13504; 117R: J23101+Ribo J+I13504; D: E. coli K-12 DH5α without plasmid; R: R0040; I: I0270; 1: Technical replicate 1; 2: Technical replicate 2; 3: Technical replicate 3; A:Biological replicate 1; B: Biological replicate 2; C: Biological replicate 3; M9: M9 liquid media; \: Blank control'SF: Sodium Fluorescein (concentration of 0, 10, 20, 40, 80, 160, 320, 640 ng/mL)
                  </p>
                  <h3>Unit</h3>
@@ Line 46: / Line 39: @@
                  <p></p>
                  <p>TR: Technical Replicate<BR>BR: Biological Replicate</p>
-                 <div style="width: 60%;margin: 0 auto;">
+                 <div style="width: 100%;margin: 0 auto;">
                      <center>https://static.igem.org/mediawiki/2015/4/4e/OUC-China-InterLab_5.png</center>
                  </div>