CytoScan™ HD Training Kit

Here is the link to the CytoScan assay user guide: https://assets.thermofisher.com/TFS-Assets/LSG/manuals/703038_cytoscan_assay_UG.pdf
In Appendix D (starting on page 67), the manual outlines specifications for compatible thermocyclers. Any thermocycler that fits those specifications will work.

Note: Use the silver or gold-plated block only, do not use an aluminum block. In order to determine which block you have, look at the serial number on the block. An "A" in the serial number indicates that the block is aluminum; an "S" indicates that it is silver. Also, an aluminum block has a honeycomb appearance between the wells, whereas a silver block is smooth.

Methods that include boiling or strong denaturants are not acceptable, because the DNA would be rendered single-stranded. For genomic DNA extraction, we recommend using either the Qiagen Gentra Puregene Kit or the 5 Prime PerfectPure DNA Blood Kit.

Our CytoScan HD Assay requires genomic DNA concentration ≥50 ng/µL. Therefore, the elution volumes for each of the kits will need to be adjusted accordingly to achieve the desired concentration.

The presence of RNA and free nucleotides can interfere with some quantitation methods using a spectrophotometer or a NanoDrop instrument. To eliminate RNA contamination, perform RNase treatment during extraction as follows:
- For QIAGEN Gentra Puregene Kit: Perform RNase treatment as recommended in the extraction kit manual prior to elution of genomic DNA.
- For 5 PRIME PerfectPure DNA Blood Kit: Use only RNase-treated purification columns for extraction of genomic DNA.

The purified genomic DNA extracted using the two methods above should meet the DNA quality specifications per the manufacturer’s kit extraction manual.
Note: Both blood and cell line sample sources have been tested for the CytoScan HD assay.

The CytoScan HD Array includes 750,000 "genotype-able" SNPs and 1.9 million non-polymorphic probes. The CytoScan HD Array covers both constitutional and cancer genes with: - ISCA constitutional coverage at 1 marker /384 bases
- Complete cancer gene coverage at 1 marker / 553 bases
- 12,000 OMIM genes at 1 marker / 659 bases
- >36,000 RefSeq genes with 1 marker / 880 bases
- Backbone (non-gene) coverage of 1 marker / 1,737 bases across genome for breakpoints

Array format: 49
Feature size: 5 µM
Time to scan per array: ~32 mins
Hyb temperature: 50 degrees C

The CytoScan HD Array uses the following QC metrics:
- SNPQC ≥15
- MAPD ≤0.25
- Waviness SD ≤0.12

The Purification Wash Buffer and Elution Buffer are not affected by being frozen at -20 degrees C and are safe to use for purification.

A subset of Y probes is used and run through a Hidden Markov Model to determine gender. Copy number of 0 or 1. 0 indicates female and 1 indicates male.

Find additional tips, troubleshooting help, and resources within our Microarray Analysis Support Center.