PCR of dCTailed cDNA

The described precautions against DNA contamination must be followed when setting up this and the following PCR (15). Pre- and post-PCR procedures and equipment are separated geographically; all solutions are stored as aliquots and only used once. Thus, in addition of a thermocycler, a flow cabinet is strongly recommended for setting up the PCR reaction. Use the corresponding thin-walled polypropylene tubes and keep them in wet ice.

1. Sterilized, deionized water.

2. 10X PCR buffer: 200 mM Tris HCl, pH 8.4, 500 mM KCl.

5. 10 mM ALK2 (5'-ATCAGCAAATTCAACCACCAG-3') or control GSP2 (for control reaction).

6. 10 mM Abridged Anchor Primer AAP (5'-GGCCACGCGTCGACTAGTAC GGGIIGGGIIGGGIIG-3').

7. 5 U/|L of Taq DNA pol (Boehringer Mannheim, Mannheim, Germany).

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