User:Karmella Haynes/Notebook/BioBrick cloning/2013/03/19

From OpenWetWare
< User:Karmella Haynes‎ | Notebook‎ | BioBrick cloning‎ | 2013‎ | 03
Revision as of 20:32, 19 March 2013 by Karmella Haynes (talk | contribs) (03/19/13)
Jump to: navigation, search
Owwnotebook icon.pngKarmella's BioBrick Cloning <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

03/19/13

  • Oligo annealing for new mammalian vector based on MV9



Oligo annealing

  • Note: made fresh 10x annealing buffer
  1. MV9+ K-XbaI-NLS-his-stop
  • K-XbaI-NLS-his-st 1
  • K-XbaI-NLS-his-st 2
  • K-XbaI-NLS-his-st 3
  • K-XbaI-NLS-his-st 4
DNA (oligos, 100 μM) 4x 3 μL ea. = 12.0
10x annealing buffer 2.0
dH2O 6.0
  20 μL

Thermal Cycling (try new thermal cycling approach)
Multigene Optimax Thermal Cycler

  • 99.9 °C, 10 min.
  • [99.9°C, temp. increment -2.0°C, 1 min.; 90.0°C, temp. increment -2.0°C, 1 min.] x35
  • 25°C, ∞

Vector Digest