Please make sure your are using the “…gene_exp.diff” file that comes from CuffLinks. There are some applications that claim to emulate CuffDiff output (e.g. Galaxy). If you are using one of these applications, please make sure the output file has all columns populated. See below for specific columns that must be present. Also, use this link to view the Cuffdiff manual.

Column number Column name Example Description
1 Tested id XLOC_000001 A unique identifier describing the transcipt, gene, primary transcript, or CDS being tested
2 gene Lypla1 The gene_name(s) or gene_id(s) being tested
3 locus chr1:4797771-4835363 Genomic coordinates for easy browsing to the genes or transcripts being tested.
4 sample 1 Liver Label (or number if no labels provided) of the first sample being tested
5 sample 2 Brain Label (or number if no labels provided) of the second sample being tested
6 Test status NOTEST Can be one of OK (test successful), NOTEST (not enough alignments for testing), LOWDATA (too complex or shallowly sequenced), HIDATA (too many fragments in locus), or FAIL, when an ill-conditioned covariance matrix or other numerical exception prevents testing.
7 FPKMx 8.01089 FPKM of the gene in sample x
8 FPKMy 8.551545 FPKM of the gene in sample y
9 log2(FPKMy/FPKMx) 0.06531 The (base 2) log of the fold change y/x
10 test stat 0.860902 The value of the test statistic used to compute significance of the observed change in FPKM
11 p value 0.389292 The uncorrected p-value of the test statistic
12 q value 0.985216 The FDR-adjusted p-value of the test statistic
13 significant no Can be either “yes” or “no”, depending on whether p is greater then the FDR after Benjamini-Hochberg correction for multiple-testing