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
(mm/dd/yy)
Current revision (17:17, 22 March 2013) (view source)
(03/19/13)
 
(7 intermediate revisions not shown.)
Line 16: Line 16:
* Note: made fresh 10x annealing buffer
* Note: made fresh 10x annealing buffer
-
# New BB 1
+
# KAH3/MV2(X) + K-XbaI-NLS-his-stop
-
* oligo 1
+
* K-XbaI-NLS-6his-st 1
-
* oligo 2
+
* K-XbaI-NLS-6his-st 2
-
* oligo 3
+
* K-XbaI-NLS-6his-st 3
-
* oligo 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
|-
|-
Line 32: Line 32:
|}
|}
-
Thermal Cycler (try new thermal cycling approach)<br>
+
Thermal Cycling (try new thermal cycling approach)<br>
Multigene Optimax Thermal Cycler
Multigene Optimax Thermal Cycler
* 99.9 °C, 10 min.
* 99.9 °C, 10 min.
-
* [99.9°C, temp. increment -2.0°C, 1 min.; 97.9°C, temp. increment -2.0°C, 1 min.] x35
+
* [99.9°C, temp. increment -2.0°C, 1 min.; 90.0°C, temp. increment -2.0°C, 1 min.] x35
* 25°C, ∞
* 25°C, ∞
 +
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->

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, ∞



Personal tools