Preparation of DIGLabeled Single Stranded RNA Probe Riboprobe by In Vitro Transcription

1. Riboprobe Gemini System II Buffers Kit (Promega, Southampton, UK) includes cold ribonucleotides, DTT, Transcription optimized buffer. Store at -20°C. NTPs go off, so kits to suit level of use should be bought.

2. T3 and T7 RNA polymerase (Promega, Southampton, UK). Store at -20°C.

3. DIG-UTP (Boehringer Mannheim, Lewes, UK). Store at -20°C.

4. RNasin Ribonuclease Inhibitor (Promega, Southampton, UK). Store at -20°C.

5. Restriction enzymes (single cut 5' and 3' of insert).

6. Phenol and chloroform.

7. 7.8 M Ammonium acetate. 8.4 M Lithium chloride.

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