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

From OpenWetWare

< User:Karmella Haynes | Notebook | BioBrick cloning | 2013 | 03(Difference between revisions)
Jump to: navigation, search
(→03/19/13)
Current revision (17:17, 22 March 2013) (view source)
(→03/19/13)
 
Line 22: Line 22:
* K-XbaI-NLS-6his-st 4
* K-XbaI-NLS-6his-st 4
-
{| class="wikitable" border="0" cellspacing="3" <!-- Oligo annealing rxn table -->
+
{| {{table}} <!-- Oligo annealing rxn table -->
| DNA (oligos, 100 μM) || 4x 3 μL ea. = 12.0
| DNA (oligos, 100 μM) || 4x 3 μL ea. = 12.0
|-
|-

Current revision

Karmella's BioBrick Cloning Main project page
Previous entry      Next entry

03/19/13

  • Oligo annealing for new mammalian vector


Oligo annealing

  • Note: made fresh 10x annealing buffer
  1. KAH3/MV2(X) + K-XbaI-NLS-his-stop
  • K-XbaI-NLS-6his-st 1
  • K-XbaI-NLS-6his-st 2
  • K-XbaI-NLS-6his-st 3
  • K-XbaI-NLS-6his-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, ∞



Retrieved from "http://openwetware.org/wiki/User:Karmella_Haynes/Notebook/BioBrick_cloning/2013/03/19"
Personal tools