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 (→Result) |
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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 | *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.