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We have also sequenced the tumor and normal genomes of one of the three patients and determined copy number changes present in the tumor genome.
The determined copy number of CSRNP mRNA was normalized to the amount of 18S rRNA in each sample (TaqMan® Ribosomal RNA control reagents (Applied Biosystems).
We therefore typed rs4300027 in the 589 samples for which we had determined copy number.
Recent studies (2, 31) used SNP arrays (SNP6.0) and determined copy numbers as single integer values.
F r1) gives a measure of how distant the determined copy numbers are from integer numbers, if r1 is chosen as representing copy number 1.
Using data from 29 Affymetrix SNP 6.0 arrays, we determined copy numbers using three programs: Partek Genomics Suite, Affymetrix Genotyping Console 2.0 and Birdsuite.
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However, this single-pot strategy requires in silico masking of the coding sequences (exonic region) of the genome to determine copy number variation, which leads to incomplete transcripts from the cell.
In order to determine copy number changes, CGH-Explorer v. 3.1b was used [16].
The Affymetrix chip determines copy number based on values of nearly 1,000,000 probes in the genome, resulting in a high probability for both type I and II errors.
We sequenced the tumor and normal genomes of one patient and used these data to determine copy number changes (CNCs) between the two samples.
For determining copy number gains and losses, the R package CGH call was used [ 16].
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