Christian Niederauer/Notebook/RacingBacteria/Stock: Difference between revisions

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=Stock=
=Bacteria=
==Bacteria==
====Master Plate====
*[[Christian Niederauer/Notebook/RacingBacteria/Stock/Master_Plate | Master Plate]] with [[Christian Niederauer/Notebook/RacingBacteria/Stock/RP437 |RP437 E.Coli]]
Created on [http://openwetware.org/wiki/Christian_Niederauer/Notebook/RacingBacteria/Calendar#12.08.2014_Reinoculate_motile_colony 12/08/2014] from liquid culture, which again was created from one motile colony of the original master plate.<br> The original master plate again was created from glycerol stock of [[Christian_Niederauer/Notebook/RacingBacterias/Stock#RP437_Strain | RP437 E. Coli]] on [http://openwetware.org/wiki/Christian_Niederauer/Notebook/RacingBacteria/Calendar#06.08.2014_Revive_Bacteria 06.08.].
* 1ml [[Christian Niederauer/Notebook/RacingBacteria/Stock/Liquid_Colony | Liquid Colony]] from [[Christian Niederauer/Notebook/RacingBacteria/Stock/Master_Plate | Master Plate]]


==Microfluidics Devices==
====Liquid Colony====
*[[Christian Niederauer/Notebook/RacingBacteria/Stock/Leveling_Petri_Dish | Leveling Petri Dish]]
*Created on 19/08/2014 ''[disappeared]''<br>
* various non- or bad-functioning racing tracks
Suspended a few colonies from [[Christian_Niederauer/Notebook/RacingBacteria/Stock#Master_Plate | Master Plate]] into 1ml of LB media.
* Mother Machine Stamp (without holes / connections)
* Original Silicon Wafer with [[Christian Niederauer/Notebook/RacingBacteria/Stock/H-Pattern | H-Pattern]]


*Created on 27/08/2014<br>
Suspended a few colonies from [[Christian_Niederauer/Notebook/RacingBacteria/Stock#Master_Plate | Master Plate]] into ~5ml of LB media.


==Material==
====RP437 Strain====
[http://openwetware.org/images/0/01/RP437_bacteria_strain.pdf Datasheet of the RP437 Strain]
 
After reviving this strain, test on soft agar plates and pick a colony that is motile and chemotactic because nonmotile variants arise in room temperature stab-cultures.
 
=Microfluidics Devices=
====Leveling Petri Dish====
'''Problem:''' Oven is not even, the PDMS stamps therefore will be to thin at one side or the fluid even leaks over the waver and the stamp will be to thin everywhere.<br>
'''Solution:''' Filling up a Petri Dish with ~20ml PDMS and curing it in the oven (lower rack, placing dish at the very back on the right wall with the marker pointing perpendicular to the backwall).
The fluid's plane will be arranged perpendicular to the gravitational force eventually.
Future petri dishes with PDMS then can be placed on top of this device.
 
====H-Pattern PDMS stamps====
{|class="wikitable"
|-
| <gallery>Image:H_pattern.jpg | close up of 20.08. stamp</gallery> || <gallery>Image:H_pattern_wholes.jpg | problem: jagged holes, glass residues in stamp </gallery>
|}
*20.08. (entire pattern)
*22.08. (pre-cut)
*27.08. (Araldite Replica II)
 
====Various non- or bad-functioning racing tracks====
 
====Mother Machine Stamp (without holes / connections)====
====Original Silicon Wafer with H-Pattern====
<gallery>Image:Silicon.jpg</gallery>
[http://openwetware.org/wiki/Image:RaBa_h_pattern.pdf Original Pattern etched on the Silicon Wafer]<br>
Track width: 2µm<br>
Track length: 362µm<br>
Number of tracks: 32
 
====Araldite Molds====
{| class='wikitable'
| <gallery>Image:RaBa_ara2.jpg | Araldite Replica II in the making </gallery> || <gallery>Image:RaBa_ara2b.jpg | Araldite Replica II</gallery>
|}
*Araldite Replica I, [http://openwetware.org/wiki/Christian_Niederauer/Notebook/RacingBacteria/Calendar#26.08.2014_ACTUAL_Araldite_Replica_from_PDMS_.26_COMSOL created on 26.08.] from H-Pattern PDMS stamp (20.08.)
*Araldite Replica II, [http://openwetware.org/wiki/Christian_Niederauer/Notebook/RacingBacteria/Calendar#26.08.2014_ACTUAL_Araldite_Replica_from_PDMS_.26_COMSOLcreated on 26.08.] from H-Pattern PDMS stamp (20.08.) with increased thickness
 
=Material=


*Coring Tool 2mm
*Coring Tool 2mm
*PDMS Chemicals
*PDMS Chemicals (Sylgard, Silicone Elasteromer KIT 184)
* Araldite Chemicals (Resinova Bondtite)

Latest revision as of 23:55, 26 August 2014

Bacteria

Master Plate

Created on 12/08/2014 from liquid culture, which again was created from one motile colony of the original master plate.
The original master plate again was created from glycerol stock of RP437 E. Coli on 06.08..

Liquid Colony

  • Created on 19/08/2014 [disappeared]

Suspended a few colonies from Master Plate into 1ml of LB media.

  • Created on 27/08/2014

Suspended a few colonies from Master Plate into ~5ml of LB media.

RP437 Strain

Datasheet of the RP437 Strain

After reviving this strain, test on soft agar plates and pick a colony that is motile and chemotactic because nonmotile variants arise in room temperature stab-cultures.

Microfluidics Devices

Leveling Petri Dish

Problem: Oven is not even, the PDMS stamps therefore will be to thin at one side or the fluid even leaks over the waver and the stamp will be to thin everywhere.
Solution: Filling up a Petri Dish with ~20ml PDMS and curing it in the oven (lower rack, placing dish at the very back on the right wall with the marker pointing perpendicular to the backwall). The fluid's plane will be arranged perpendicular to the gravitational force eventually. Future petri dishes with PDMS then can be placed on top of this device.

H-Pattern PDMS stamps

  • 20.08. (entire pattern)
  • 22.08. (pre-cut)
  • 27.08. (Araldite Replica II)

Various non- or bad-functioning racing tracks

Mother Machine Stamp (without holes / connections)

Original Silicon Wafer with H-Pattern

Original Pattern etched on the Silicon Wafer
Track width: 2µm
Track length: 362µm
Number of tracks: 32

Araldite Molds

  • Araldite Replica I, created on 26.08. from H-Pattern PDMS stamp (20.08.)
  • Araldite Replica II, on 26.08. from H-Pattern PDMS stamp (20.08.) with increased thickness

Material

  • Coring Tool 2mm
  • PDMS Chemicals (Sylgard, Silicone Elasteromer KIT 184)
  • Araldite Chemicals (Resinova Bondtite)