Hosuk:LabNotesMDA/2013-7-18: Difference between revisions

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>Hosuki78
(Created page with "*LabNote ===MDA with PGP1F, 8th, Dev v9.1=== *Use new device (v9.1) ====KOH concentration for (ALS+Cell) mix==== *I...")
 
>Hosuki78
 
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====KOH concentration for (ALS+Cell) mix====
====Experiment====
=====KOH concentration for (ALS+Cell) mix=====
*In 20uL reaction, the final concentration of KOH in ALS while denaturing is '''240mM'''.
*In 20uL reaction, the final concentration of KOH in ALS while denaturing is '''240mM'''.
*In the prev version,269mM of KOH in ALS buffer has used ([[Hosuk:LabNotesMDA/2013-7-2|v7.3 ring mixer device]]).
*In the v7.3 version, '''269mM''' of KOH in ALS buffer has used ([[Hosuk:LabNotesMDA/2013-7-2|v7.3 ring mixer device]]).
*For this v9.1 device, 286mM of KOH in ALS buffer is used (Cell : (ALS-Cell) = 1 : 5.247).
*For this v9.1 device, '''286mM''' of KOH in ALS buffer is used (Cell : (ALS-Cell) = 1 : 5.247).




*[[File:ALS_Recipe.png|600px]]
*[[File:ALS_Recipe.png|600px]]
*Start : 8:30pm 07/17, End : 9:00am 07/18 --> about 13hr reaction
*qPCR run for verification
====Result====
*Amount of amplicons were good. (output channel 22 was clogged.)
*Some of sample could be lost because of back flow as soon as opening valve
*So the left 4 chamber were not filled well.
[[File:Backflow_after_opening_valve_2013-07-18.png|400px]]
*I didn't run MDA in ALS chamber, but the reaction will be done in ALS chamber as well for the next experiment.
*Also I'll try to push the (ALS+Cell) solution into compartment with '''H2O''' to prevent sample loss like this.
*[[File:qPCR_Result_MDAv9.1_2013-07-18_fig1.png|650px]]
*[[File:qPCR_Result_MDAv9.1_2013-07-18_fig2_Table.png|400px]]
*[[File:qPCR_Result_MDAv9.1_2013-07-18_fig3_History.png|600px]]

Latest revision as of 18:44, 26 July 2013


MDA with PGP1F, 8th, Dev v9.1[edit]

  • Use new device (v9.1)


Experiment[edit]

KOH concentration for (ALS+Cell) mix[edit]
  • In 20uL reaction, the final concentration of KOH in ALS while denaturing is 240mM.
  • In the v7.3 version, 269mM of KOH in ALS buffer has used (v7.3 ring mixer device).
  • For this v9.1 device, 286mM of KOH in ALS buffer is used (Cell : (ALS-Cell) = 1 : 5.247).



  • Start : 8:30pm 07/17, End : 9:00am 07/18 --> about 13hr reaction
  • qPCR run for verification


Result[edit]

  • Amount of amplicons were good. (output channel 22 was clogged.)
  • Some of sample could be lost because of back flow as soon as opening valve
  • So the left 4 chamber were not filled well.

File:Backflow after opening valve 2013-07-18.png


  • I didn't run MDA in ALS chamber, but the reaction will be done in ALS chamber as well for the next experiment.
  • Also I'll try to push the (ALS+Cell) solution into compartment with H2O to prevent sample loss like this.