Hosuk:LabNotes/2013-6-10: Difference between revisions

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*Phi29 enzyme from '''Enzymatics''' is used.
*Phi29 enzyme from '''Enzymatics''' is used.
*Actually Jay likes to use (or trusts) Enzymatics, so almost all reagents are from Enzymatics.
*Actually Jay likes to use (or trusts) Enzymatics, so almost all reagents are from Enzymatics.
*The concentration of Phi29 of Enzymatics that Jonathan has used is '''100U/uL''' which is '''High concentration one''' from Enzymatics, and this concentration is the same as low concentration one from Epicentre.
**'''1uL of phi29 in 200uL total mix'''
*RNase H step after RT is critical step, so do this carefully.
*RNase H step after RT is critical step, so do this carefully.
**'''10uL of RNase H''' +
**'''2uL of Riboshredder''' +
**'''20uL of 10x Buffer''' +
**168uL of H2O
*Try HeLa cells unless I get rolonies with PGP1F.
*Try HeLa cells unless I get rolonies with PGP1F.



Revision as of 19:54, 10 June 2013

Training Day 1
  • Some of important things
  • Phi29 enzyme from Enzymatics is used.
  • Actually Jay likes to use (or trusts) Enzymatics, so almost all reagents are from Enzymatics.
  • The concentration of Phi29 of Enzymatics that Jonathan has used is 100U/uL which is High concentration one from Enzymatics, and this concentration is the same as low concentration one from Epicentre.
    • 1uL of phi29 in 200uL total mix


  • RNase H step after RT is critical step, so do this carefully.
    • 10uL of RNase H +
    • 2uL of Riboshredder +
    • 20uL of 10x Buffer +
    • 168uL of H2O


  • Try HeLa cells unless I get rolonies with PGP1F.


  • The sample (that I've got some signal at 05/29) doesn't seem to have actual rolonies,
  • There are no dots, just too much even signal distribution.