Proteinase K, 100 mg E76230Y
SpectralChip™ 2600 BAC Array, 5 pack SC2600-5PK*
Constitutional Chip™ 1.0 BAC Array, 5 pack CC1.0-5PK*
*GE Healthcare is the exclusive distributor of Spectral Genomics products outside the United States and Canada.
Other materials required
FFPE human liver tissue (Asterand)
Chelex-100 resin (Bio-Rad)
DOP-PCR primers (Integrated DNA Technologies)
GenePix™ 4000B Microarray Scanner (Axon Instruments)
GenePix Pro 3.1 Microarray Image Analysis Software (Axon Instruments)
Thermal cycler (MJ Research)
Quant-iT™ PicoGreen™ dsDNA Assay Kit (Invitrogen)
HyperLadder I DNA ladder (BioLine)
Dpn II restriction endonuclease (New England BioLabs)
DNA extraction from FFPE tissue slides
Genomic DNA was isolated using Chelex-100 from a 20-µm section of FFPE liver tissue. The detailed protocol is given below. DNA was quantitated using PicoGreen dsDNA quantitation reagent.
1. Add 2 µl of 10 mg/ml Proteinase K to each tube (final concentration 200 µg/ml).
2. Incubate at 55 °C for 3 h. Agitate gently every hour.
3. Add 100 µl of 5% Chelex-100 in Tris-EDTA.
4. Heat to 99 °C for 10 min.
5. Shake gently and spin at 10,500 x g for 15 min (while still hot).
6. Place on ice and remove any hard