Hosuk:LabNotes/2014-9-30

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Tissue Section : fixing test[edit]

  • Continued from 2014-09-29
  • RCA : start at 2014-09-30, 1pm~ (incubate for overnight)


ppCOL1A1 + Suppressor test[edit]

  • Sample : 4 dishes PGP1F, 1st Rolonies made at 2014-07-17
  • Check 1st rolonies of all 5 dishes
  • S1, S4 --> ppCOL1A1 + suppCOL1A1
  • S2, S5 --> ppCOL1A1 only
  • Add suppressor 10 times more than ppCOL1A1


File:Fig MIP PGP1F RCA 2014-07-17 1stRolony 3.png


Procedure[edit]

  • ppCOL1A1 + supp_COL1A1 : S1, S4
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
100uM ppCOL1A1 2
100uM Supp_COL1A1 20
H2O 58
Total 100


  • ppCOL1A1 : S2, S5
Components Volume
5 U/ul Ampligase 10
10X Ampligase Buffer 10
100uM ppCOL1A1 2
100uM Supp_COL1A1 0
H2O 78
Total 100


  • Incubate at 37C for 30min
  • Incubate at 60C for 24hrs