tissue: Kidney transplant biopsy i-ifta grade: 1 total i score: 1 cadi score: 3 time point: 06 months
Treatment protocol
Tacrolimus-Mycophenolate mofetil
Extracted molecule
total RNA
Extraction protocol
each kidney allograft biopsy (n=120), a separate biopsy core was placed in RNAlater (Ambion Inc, Austin, TX, USA) and stored at –20°C until RNA extraction. Total RNA was extracted from each biopsy using TRIzol Reagent (Invitrogen, Carlsbad, CA, USA).
Label
biotin
Label protocol
biotin labeling using the OvationTM Biotin System (NuGEN Technologies, San Carlos, CA, USA)
Hybridization protocol
The cDNA fragments of all 120 biopsies were hybridized onto Affymetrix GeneChip® Human Genome U133 Plus 2.0 Arrays comprising more than 54,000 probe sets, covering more than 47,000 transcripts and variants, including 38,500 well-characterized human genes (Affymetrix Inc., Santa Clara, CA, USA).
Scan protocol
The microarrays were scanned using GeneChip Scanner 3000 (Affymetrix Inc., Santa Clara, CA, USA).
For processing and normalization of the scanned images, dChip 2006 software was used, with perfect match [PM]/mismatch [MM] difference modeling and invariant set normalization
