Preparation of siRNA

1. The 21-nt RNA oligonucleotides are chemically synthesized and purified with high-performance liquid chromatography (HPLC) by MWG (Germany).

2. Synthetic oligonucleotides are then deprotected using the methods and solution supplied by MWG; they are then dissolved in DNase/RNase-free dH2O at 25 |jM concentration.

3. The single-strand RNA oligonucleotides are aliquoted and stored at -80°C.

Fig. 2. Sequences of E6, E7, and control siRNAs. (Reproduced with permission from ref. 11.)

4. Anneal the siRNAs as follows:

Sense RNA oligonucleotides (25 pM)

Antisense RNA oligonucleotides (25 pM)

10X Annealing buffer ddH2O


Volume (pL) 40 40 10 10 100

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