Difference between revisions of "Team:Utrecht/testpage"

Line 287: Line 287:
  
 
<h5>Calibration 2 </h5>
 
<h5>Calibration 2 </h5>
 +
<ul>
 
<li>MS= 200 ul Microsphere Stock Solution</li>
 
<li>MS= 200 ul Microsphere Stock Solution</li>
 
<li>dd= 100 uL ddH20</li>
 
<li>dd= 100 uL ddH20</li>
 
<li>green= serial dilution was performed with a micropipet from E1,F1,G1,H1 - E11,F11,G11,H11 by a volume of 100uL.            Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.</li>  
 
<li>green= serial dilution was performed with a micropipet from E1,F1,G1,H1 - E11,F11,G11,H11 by a volume of 100uL.            Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.</li>  
 
<li>Measurement: Abs600, re-mix befor putting in plate reader and prevent bubbles, path length correction off</li>
 
<li>Measurement: Abs600, re-mix befor putting in plate reader and prevent bubbles, path length correction off</li>
 +
</ul>
  
 +
<h5>Calibration 3</h5>
 +
<ul>
 +
<li>1xFC= 200 mL 1xFC (100uL 10x fluorescein + 900ul 1x PBS pH 7.4, tube was covered with foil</li>
 +
<li>P= 100 uL 1x PBS pH 7.4</li>
 +
<li>green= serial dilution was performed with a micropipet from A1,B1,C1,D1 - A11,B11,C11,D11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged. </li>
 +
<li>Measurement: FL, 530nm/30nm bandpass, 25-30nm with recommened excitation of 485nm, emission 520-530nm of the filter. Path length correction was turned off</li>
 +
</ul>
  
 
<table class="InterlabTable">
 
<table class="InterlabTable">
Line 305: Line 314:
 
<tr><td>H</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr>
 
<tr><td>H</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr>
 
</table>
 
</table>
−
 
−
<h5>Calibration 3</h5>
 
−
<li>1xFC= 200 mL 1xFC (100uL 10x fluorescein + 900ul 1x PBS pH 7.4, tube was covered with foil</li>
 
−
<li>P= 100 uL 1x PBS pH 7.4</li>
 
−
<li>green= serial dilution was performed with a micropipet from A1,B1,C1,D1 - A11,B11,C11,D11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged. </li>
 
−
<li>Measurement: FL, 530nm/30nm bandpass, 25-30nm with recommened excitation of 485nm, emission 520-530nm of the filter. Path length correction was turned off</li>
 
−
 
  
 
07/08/18 (afternoon)
 
07/08/18 (afternoon)

Revision as of 21:01, 6 October 2018


PLACEHOLDER

Interlab

August
September

Receptor Assay

July
August
September

BRET Assay

July
August
September

Methylation Assay

July
August
September
Mo
Tu
We
Th
Fr
Sa
Su

Morning

Performed Calibration 1, 2, and 3 according to protocol. All measurements were performed using the plate reader of Seino.

Table 1. content 96 wells plate cal.1 and cal.2
123456789101112
ALdd
BLdd
CLdd
DLdd
E MSdddddddddddd dddddddddd
FMSdddddddddddd dddddddddd
G MSdddddddddddddddddddddd
H MSdddddddddddddddddddddd
Calibration 1
  • L= 100 uL Ludox CL-X (stored at 4C)
  • dd= 100 uL ddH20
  • Measurement: Abs600, turn off pathlength correction
Calibration 2
  • MS= 200 ul Microsphere Stock Solution
  • dd= 100 uL ddH20
  • green= serial dilution was performed with a micropipet from E1,F1,G1,H1 - E11,F11,G11,H11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
  • Measurement: Abs600, re-mix befor putting in plate reader and prevent bubbles, path length correction off
Calibration 3
  • 1xFC= 200 mL 1xFC (100uL 10x fluorescein + 900ul 1x PBS pH 7.4, tube was covered with foil
  • P= 100 uL 1x PBS pH 7.4
  • green= serial dilution was performed with a micropipet from A1,B1,C1,D1 - A11,B11,C11,D11 by a volume of 100uL. Before every transfer solution was pipetted up and down 3x, after every transfer tips were discharged.
  • Measurement: FL, 530nm/30nm bandpass, 25-30nm with recommened excitation of 485nm, emission 520-530nm of the filter. Path length correction was turned off
Table 2. content 96 wells plate cal.3
123456789101112
A1xFCPPPPPPPPPPP
B1xFCPPPPPPPPPPP
C1xFCPPPPPPPPPPP
D1xFCPPPPPPPPPPP
E
F
G
H
07/08/18 (afternoon) LBC plates were made according to the protocol used on the wall 250ml LB 2x added to melted 250 ml WA 2x using a microwave 0.5ml was added to final solution plates were dried in 37C incubator Transformation device 3 + negative control interlab study Device 3 (number 5) showed a low GFP expression, so it was tried to re-preform the tranformation. Negative control of the interlab (number 1) was not performed last time due to lack of LBC plates so was also performed. protocol used: competent cells (DH5-a) was put on ice for 20 min 50ul of competent cells were transferred to inoculation tube, one for each separate DNA sample 2ul DNA (number 5 or 1) was added 30 min on ice 42 C heatshock for 45 sec coldshock on ice 5 min 500uL SOB was added to each of the tubes 1h incubated in 37C shaker plated on LBC plates and incubated overnight at 37 C 100ul directly (with resuspending) centrifugation at full speed for 30 sec 300ul was removed pellet was resuspended in remaining 100ul 100ul was plated onto LBC plate
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