Huang: DH5alpha transformation

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(New page: 1. Set tubes of frozen cells on ice and let thaw very slowly 2. Add 100ul thawed cells to new eppendorf 3. Add 1-2ul DNA to cells 4. Incubate on ice for 20 min 5. Heat shock in 42C water b...)
Current revision (19:49, 1 April 2009) (view source)
 
Line 1: Line 1:
-
1. Set tubes of frozen cells on ice and let thaw very slowly
+
==Overview==
-
2. Add 100ul thawed cells to new eppendorf
+
 
-
3. Add 1-2ul DNA to cells
+
This protocol allows competent cell transformation.
-
4. Incubate on ice for 20 min
+
 
-
5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
+
==Materials==
-
6. Incubate on ice 2 min
+
*DH5alpha competent cells
-
7. Add 900ul growth medium (LB) and shake 1 hour at 37C
+
*LB
-
8. Plate about 200ul and grow overnight
+
*LB + antibiotic plates
 +
*Plasmid DNA or DNA to transform the cells with
 +
 
 +
==Procedure==
 +
*1. Set tubes of frozen cells on ice and let thaw very slowly
 +
*2. Add 100ul thawed cells to new eppendorf
 +
*3. Add 1-2ul DNA to cells
 +
*4. Incubate on ice for 20 min
 +
*5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
 +
*6. Incubate on ice 2 min
 +
*7. Add 900ul growth medium (LB) and shake 1 hour at 37C
 +
*8. Plate about 200ul and grow overnight
 +
 
 +
 
 +
==Contact==
 +
*[[User:Carolina Tropini|Carolina Tropini]]

Current revision

Contents

Overview

This protocol allows competent cell transformation.

Materials

  • DH5alpha competent cells
  • LB
  • LB + antibiotic plates
  • Plasmid DNA or DNA to transform the cells with

Procedure

  • 1. Set tubes of frozen cells on ice and let thaw very slowly
  • 2. Add 100ul thawed cells to new eppendorf
  • 3. Add 1-2ul DNA to cells
  • 4. Incubate on ice for 20 min
  • 5. Heat shock in 42C water bath for one min (make sure water bath is at exactly 42C)
  • 6. Incubate on ice 2 min
  • 7. Add 900ul growth medium (LB) and shake 1 hour at 37C
  • 8. Plate about 200ul and grow overnight


Contact

Personal tools