Haynes Lab:Notebook/Engineering PC-TFs/2013/10/19: Difference between revisions

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Line 14: Line 14:
| align="center" style="background:#efefef;"|'''Control'''
| align="center" style="background:#efefef;"|'''Control'''
|-
|-
| DNA Insert||1.4||1.8
| DNA Insert||0.8||0
|-
|-
| DNA Vector (Kozak)||0.6||0.6
| DNA Vector (MV9)||2.4||2.4
|-
|-
| T4 Ligase||1||1
| T4 Ligase||1||1
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| Lign Buffer (2x)||5||5
| Lign Buffer (2x)||5||5
|-
|-
| dH2O||2.0||1.6
| dH<sub>2</sub>O ||0.8||1.6
|-
|-
| '''Total'''||10 µL||10 µL
| '''Total'''||10 µL||10 µL

Revision as of 14:08, 20 October 2013



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October 19, 2013

Ligation of CMV + MV9

1 Control
DNA Insert 0.8 0
DNA Vector (MV9) 2.4 2.4
T4 Ligase 1 1
Lign Buffer (2x) 5 5
dH2O 0.8 1.6
Total 10 µL 10 µL


Bacterial Transformation of CMV Promoter + MV9

  • Warm 100 μg/mL Amp agar plate at 37°C
  • Thaw fresh tube of DH5α Turbo cells on ice
  • Add 30 μL of DH5α Turbo Cells to DNA + dH2O
  • Incubate cells + DNA on ice for 5 minutes.
  • Label pre-warmed plate.
  • Transfer cells + DNA onto agar.
  • Add 8-10 glass beads, shake, then discard beads.

Incubate plates at 37°C overnight and retrieve next day.



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