Question: RSEM: The reference contains no transcripts!
0
gravatar for Dylan Wang
11 months ago by
Dylan Wang40
Harbin
Dylan Wang40 wrote:

I want to use RSEM tool to finish my job. However, I got some question. Here is the code:

rsem-prepare-reference --gtf /home/dwang/Data/GRCh/Homo_sapiens.GRCh37.68.gtf --star -p 2 ~/Data/UCSC/hg19.fa /Data/Results

There are some errors. I do not know why.

Warning: 197611 transcripts are failed to extract because their chromosome sequences are absent.
0 transcripts are extracted.
The reference contains no transcripts!

I downloaded the annotation data from "ensembl" and the reference genome from UCSC.

sequencing rna-seq alignment • 644 views
ADD COMMENTlink modified 11 months ago by WouterDeCoster42k • written 11 months ago by Dylan Wang40

I edited your title to make it more specific about what you are asking.

ADD REPLYlink written 11 months ago by WouterDeCoster42k

Django : If an answer was helpful, you should upvote it; if the answer resolved your question, you should mark it as accepted. You can accept more than one if they work.
Upvote|Bookmark|Accept

Please do the same for your previous posts as well.

ADD REPLYlink written 11 months ago by genomax73k
2
gravatar for WouterDeCoster
11 months ago by
Belgium
WouterDeCoster42k wrote:

I downloaded the annotation data from "ensembl" and the reference genome from UCSC.

Well there's your problem. Use annotation from the same source as your reference genome. Your chromosome identifiers have to match.

ADD COMMENTlink written 11 months ago by WouterDeCoster42k

Thank you for your answer! I downloaded some SRA data from NCBI. I read its introduction:

Data processing Basecalls performed using bcl2fastq (v2-2.19).
Reads were aligned to hg19 reference using STAR (v2.5.2).
Transcripts were quantified using RSEM (v1.2.30) and the GRCh37.68 gtf annotation file.
Genome_build: hg19 (GRCh37)
Supplementary_files_format_and_content: *_21gene_expression.txt: Normalized expression values provided in a log2 scale.

What is the difference between hg19 and GRCh37?

ADD REPLYlink written 11 months ago by Dylan Wang40

Mainly the chromosome identifiers. The sequence is equal.

ADD REPLYlink modified 11 months ago • written 11 months ago by WouterDeCoster42k

Yep. Ensembl mark chromosomes as "1","2" etc while UCSC marks them as "chr1", "chr2" etc.

ADD REPLYlink written 11 months ago by kristoffer.vittingseerup2.5k
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