Haynes Lab:Notebook/Engineering PC-TFs/2013/03/05

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(Summary)
(Summary)
Line 20: Line 20:
* BL09
* BL09
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''B)Follow digest procedure:''
+
<font size=3>''B)Follow digest procedure:''
   DNA          20.0 µL
   DNA          20.0 µL
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   dH2O        5.0  uL
   dH2O        5.0  uL
              
              
-
               = 30.0 uL
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               = 30.0 uL </font>
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[[Image:gel3_5_2013_cutout.jpg|200px|Stock digest 3/5/2013]]
[[Image:gel3_5_2013_cutout.jpg|200px|Stock digest 3/5/2013]]
<br>
<br>
-
<font size=4>''Transformation of hPCD, fshPCD, flyPCD'' <font size=4>
+
<font size=4>''Transformation of hPCD, fshPCD, flyPCD''</font>
<br>
<br>
*Warm 100 μg/mL Amp agar plates at 37 °C
*Warm 100 μg/mL Amp agar plates at 37 °C

Revision as of 22:19, 5 March 2013

Engineering PC-TFs Main project page
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Summary

  • Procedure

A) Get biobricks and enzymes from freezer.

SpeI, PstI, XbaI

The following list of parts are:


  • BL01
  • BL05
  • BL09

B)Follow digest procedure:

  DNA          20.0 µL
  Enzyme 1     1.0  µL
  Enzyme 2     1.0  µL
  10x buffer   3.0  µL
  dH2O         5.0  uL
            
             = 30.0 uL 


Digest Order:

Biobrick Restriction Enzymes

  • 1. BL01 XbaI/PstI
  • 2. BL05 XbaI/PstI
  • 3. BL09 XbaI/PstI


Stock digest 3/5/2013 Stock digest 3/5/2013
Transformation of hPCD, fshPCD, flyPCD

  • Warm 100 μg/mL Amp agar plates at 37 °C
  • Thaw fresh tube of DH5α Turbo cells on ice
  • Add 30 μL of DH5α Turbo cells to DNA.
  • Incubate cells + DNA on ice for 5 min.
  • Label pre-warmed plates
  • Transfer cells + DNA onto agar
  • Add 10 - 15 sterile glass beads, shake, discard beads
  • Incubate plates at 37 °C overnight


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