Hosuk:LabNotes/2014-5-7: Difference between revisions
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>Hosuki78 (Created page with "*LabNote ===Test : +/- a-dUTP, and long RT incubation at 4C=== ====1st Try==== *8 MatTek dishes - PGP1F (05/06) **10% Formalin for 15min at RT(25C), **0.2...") |
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===Test : +/- a-dUTP, and long RT incubation at 4C=== | ===Test : +/- a-dUTP, and long RT incubation at 4C=== | ||
*Continued from [[Hosuk:LabNotes/2014-5-6|05/06]] | |||
* | |||
====Procedure==== | ====Procedure==== | ||
* | *05/06 | ||
*#10% Formalin for 15min at RT(25C) --> done at CERC | *#10% Formalin for 15min at RT(25C) --> done at CERC | ||
*#Wash with PBS 3 times --> done at CERC | *#Wash with PBS 3 times --> done at CERC | ||
Line 31: | Line 14: | ||
*#Aspirate 0.25% Triton X-100 in DEPC-PBS and wash with PBS 3 times | *#Aspirate 0.25% Triton X-100 in DEPC-PBS and wash with PBS 3 times | ||
*#Add RT mix and put in 4C, incubate for 15min. (1hr. for other 2 sample of target specific primers) | *#Add RT mix and put in 4C, incubate for 15min. (1hr. for other 2 sample of target specific primers) | ||
*#Move samples to 37C and incubate for 12hr. | |||
*05/07 | |||
*#Wash with PBS once | |||
*#Prepare 18uL of BS(PEG)9 in 782uL PBS (for 4 dishes, volume per each dish is 200uL) | |||
*add 200uL BS(PEG)9 mix, incubate for 1hr at RT | |||
* |
Revision as of 17:53, 7 May 2014
Test : +/- a-dUTP, and long RT incubation at 4C
- Continued from 05/06
Procedure
- 05/06
- 10% Formalin for 15min at RT(25C) --> done at CERC
- Wash with PBS 3 times --> done at CERC
- Move samples to the lab at PFBH
- Add 0.25% Triton X-100 in DEPC-PBS and incubate for 10min at room temp.
- mean while prepare RT mix and put the mix on an ice
- Aspirate 0.25% Triton X-100 in DEPC-PBS and wash with PBS 3 times
- Add RT mix and put in 4C, incubate for 15min. (1hr. for other 2 sample of target specific primers)
- Move samples to 37C and incubate for 12hr.
- 05/07
- Wash with PBS once
- Prepare 18uL of BS(PEG)9 in 782uL PBS (for 4 dishes, volume per each dish is 200uL)
- add 200uL BS(PEG)9 mix, incubate for 1hr at RT