User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/20

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.pngKarmella's BioBrick Cloning Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

05/20/15

  • Cas-tone - Phusion PCR for H3



Cas-tone - Phusion PCR for H3

  • Phusion PCR-amplify ORFs
  1. mEmerald-H3-23, H3_1 f1 / H3_40 r1
  2. mEmerald-H3-23, H3_1 f1 / H3_60 r1
  3. mEmerald-H3-23, H3_1 f1 / H3_80 r1
  4. mEmerald-H3-23, H3_1 f1 / H3_116 r1
  5. mEmerald-H3-23, H3_1 f1 / H3_136 r1
Reagent Vol Mix (x6) Expected:
1. H3-1-40 = 129
2. H3-1-60 = 189
3. H3-1-80 = 250
4. H3-1-116 = 373
5. H3-1-136 = 433
Hover name
5 μL/lane; 1% agarose; Ladder
Plasmid DNA 0.5 3.0
10 μM H3_1 f1 1.0 6.0
10 μM r primer 1.0 ---
10 mM dNTPs 1.0 6.0
5x HF buffer 10.0 60.0
Phusion Pol. 0.5 3.0
dH2O 36.0 216.0
  50.0 μL

Thermal cycler: Bio-Rad - Phusion

  • 98°C 3 min.
  • 30x[98°C, 10 sec; 70°C 30 sec; 72°C 30 sec]
  • 72°C 3 min.
  • 4°C, ∞