Hosuk:LabNotes/2014-3-13: Difference between revisions
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>Hosuki78 (Created page with "*LabNote ===Mutation Detection with Mixed with PGP1F and NCIH1975 : Ampligase, 2nd RCA=== *Continied from 03/06 *8 Ampligase...") |
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====Experiment==== | |||
*8 Ampligase reaction, | *8 Ampligase reaction, | ||
*4 NTC(no Ampligase emzyme) | *4 NTC(no Ampligase emzyme) | ||
[[File:Mix_PGP1F_NCIH1975_24wellPlates_AmpligaseMap.png| | [[File:Mix_PGP1F_NCIH1975_24wellPlates_AmpligaseMap.png|300px]] | ||
Line 16: | Line 17: | ||
*#Run RCA reaction for 24hr. (03/14 ~ 03/15) | *#Run RCA reaction for 24hr. (03/14 ~ 03/15) | ||
*#Anneal dcProbes, and Image | *#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|500px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well04.png|800px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well06.png|800px]] | |||
[[File:Mix_PGP1F_NCIH1975_Well10.png|800px]] |
Latest revision as of 22:42, 14 March 2014
Mutation Detection with Mixed with PGP1F and NCIH1975 : Ampligase, 2nd RCA[edit]
- Continied from 03/06
Experiment[edit]
- 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[edit]
- 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