Hosuk:LabNotes/2013-6-10: Difference between revisions
Jump to navigation
Jump to search
>Hosuki78 No edit summary |
>Hosuki78 No edit summary |
||
Line 3: | Line 3: | ||
*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.