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10.0 years ago
512788522
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20
After running TopHat, I hava run the following command directly(because of no replicates for each condition, i.e normal versus mutation):
cuffdiff -p 8 \
-o ./cuffdiff_out \
-L normal,mut \
-u ./genes.gtf \
./tophat/accepted_hits.bam \
./tophat_mut/accepted_hits.bam
In addition, I also have performed the differential expression analysis in the following way:
cuffmerge -g ./genes.gtf \
-p 8 \
-o ./cuffmerge_out \
-s ./ucsc.hg19.fa assembly.txt
cuffdiff -p 8 \
-o ./diff_out \
-L normal,mut \
-u ./cuffmerge_out/merged.gtf \
./tophat/accepted_hits.bam \
./tophat_mut/accepted_hits.bam
There was great difference in their output. In the first way, the gene_exp.diff
looked like:
test_id gene_id gene locus sample_1 sample_2 status value_1 value_2 log2(fold_change) test_stat p_value q_value significant
A1BG A1BG A1BG chr19:58858171-58874214 normal mut OK 33.5662 14.8501 -1.17654 -0.611634 0.50845 0.867476 no
A1BG-AS1 A1BG-AS1 A1BG-AS1 chr19:58858171-58874214 normal mut OK 1.17516 1.66153 0.499655 0.071299 0.84635 0.94951 no
A1CF A1CF A1CF chr10:52559168-52645435 normal mut NOTEST 0 0 0 0 11 no
but in the second way, it looked like:
test_id gene_id gene locus sample_1 sample_2 status value_1 value_2 log2(fold_change) test_stat p_value q_value significant
XLOC_000001 XLOC_000001 DDX11L1 chr1:11873-29370 normal mut NOTEST 0 0 00 1 1 no
XLOC_000002 XLOC_000002 OR4F5 chr1:69090-70008 normal mut NOTEST 0 0 00 1 1 no
XLOC_000003 XLOC_000003 LOC100132062,LOC100133331 chr1:323891-328581 normal mut NOTEST 0 0 0 0 1 1 no