TALENs by Ralf Kühn Wolfgang Wurst & Benedikt Wefers

Author:Ralf Kühn, Wolfgang Wurst & Benedikt Wefers
Language: eng
Format: epub
Publisher: Springer New York, New York, NY

2.2 Equipment

1.Binocular microscope.

2.Micromanipulator and microinjection system.

3.Tungsten needle.

4.Glass capillary.

2.3 Reagents

1.Plasmids from Golden Gate TALEN and TAL Effector Kit 2.0 (Addgene).

2.pBlue-TAL vector (plasmid 49401, Addgene).

3.Competent E. coli cells.

4.Proteinase K (0.2 mg/ml) in a buffer containing 0.5 % SDS (w/v).

5.Restriction enzymes BsaI, Esp3I, and Xba I.

6.T4 DNA Ligase (400 units/μl).

7.Ligation/restriction buffer 10 × (300 mM Tris–HCl, pH 7.8, 100 mM MgCl2, 100 mM dithiothreitol, 10 mM ATP).

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