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...")
 
>Hosuki78
mNo edit summary
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*4 NTC(no Ampligase emzyme)
*4 NTC(no Ampligase emzyme)


[[File:Mix_PGP1F_NCIH1975_24wellPlates_AmpligaseMap.png|400px]]
[[File:Mix_PGP1F_NCIH1975_24wellPlates_AmpligaseMap.png|300px]]




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*#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

Revision as of 18:08, 13 March 2014

Mutation Detection with Mixed with PGP1F and NCIH1975 : Ampligase, 2nd RCA


  • 8 Ampligase reaction,
  • 4 NTC(no Ampligase emzyme)

File:Mix PGP1F NCIH1975 24wellPlates AmpligaseMap.png


  • Procedure
    1. Make Ampligase mix, add
    2. Incubate at 45C for 24hr. (03/13 ~ 03/14)
    3. Run RCA reaction for 24hr. (03/14 ~ 03/15)
    4. 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