Hosuk:LabNotes/2014-3-13: Difference between revisions
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====Experiment==== | |||
*8 Ampligase reaction, | *8 Ampligase reaction, | ||
*4 NTC(no Ampligase emzyme) | *4 NTC(no Ampligase emzyme) | ||
Line 36: | Line 37: | ||
**ppEGRF_L858R_MU 2uL | **ppEGRF_L858R_MU 2uL | ||
**H2O 172 uL | **H2O 172 uL | ||
====Result : 1st Rolonies==== | |||
*Number of rolonies from most of wells are not bad. | |||
*PGP1F has more 1st rolonies than cancer cell has. | |||
**I think main reason is the size of PGP1F is much more than cancer cell's, which make a big different. | |||
**That means the density per '''area''' of rolonies seems similar at each cell type. | |||
[[File:Mix_PGP1F_NCIH1975_Counts.png|300px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well04.png|300px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well06.png|300px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well10.png|300px]] |
Revision as of 22:41, 14 March 2014
Mutation Detection with Mixed with PGP1F and NCIH1975 : Ampligase, 2nd RCA
- Continied from 03/06
Experiment
- 8 Ampligase reaction,
- 4 NTC(no Ampligase emzyme)
File:Mix PGP1F NCIH1975 24wellPlates AmpligaseMap.png
- Procedure
- Make Ampligase mix, add
- Incubate at 45C for 24hr. (03/13 ~ 03/14)
- Run RCA reaction for 24hr. (03/14 ~ 03/15)
- Anneal dcProbes, and Image
- Ampligase Mix
- 10x Buffer 20uL
- Ampligase (5U/uL) 8uL
- ppEGRF_T790M_WT 2uL
- ppEGRF_T790M_MU 2uL
- ppEGRF_L858R_WT 2uL
- ppEGRF_L858R_MU 2uL
- H2O 164 uL
- NTC Mix
- 10x Buffer 20uL
- Ampligase (5U/uL) 0uL
- ppEGRF_T790M_WT 2uL
- ppEGRF_T790M_MU 2uL
- ppEGRF_L858R_WT 2uL
- ppEGRF_L858R_MU 2uL
- H2O 172 uL
Result : 1st Rolonies
- Number of rolonies from most of wells are not bad.
- PGP1F has more 1st rolonies than cancer cell has.
- I think main reason is the size of PGP1F is much more than cancer cell's, which make a big different.
- That means the density per area of rolonies seems similar at each cell type.
File:Mix PGP1F NCIH1975 Counts.png
File:Mix PGP1F NCIH1975 Well04.png