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我试图使用 STAR 索引为受精后 99,50 小时 (99H50) 的突变库生成基因组索引，并带有 Lawson 实验室的注释。我使用的代码如下：

module load STAR; STAR --runThreadN 10 --runMode genomeGenerate --genomeDir /gpfs/ysm/scratch60/polimanti/ag2646/99H50_new_annotation/z10starindex75/ --genomeFastaFiles /gpfs/ysm/scratch60/polimanti/ag2646/Lawsonreference/genome.fa --sjdbGTFfile /gpfs/ysm/scratch60/polimanti/ag2646/Lawsonreference/genes.gtf --sjdbOverhang 75
the batch script used to submit the job for creation of such indices is 
dsq --job-file z10starindex75.txt --job-name z10starindex75 -c 10 --mem=100G -t 10:00:00 --mail-type=ALL --mail-user=aranyak.goswami@yale.edu

我试图在我的 HPC 集群上运行这段代码，它给我一个错误，如下所示：

Jan 22 22:41:39 ..... started STAR run
Jan 22 22:41:39 ... starting to generate Genome files
Jan 22 22:42:04 ... starting to sort Suffix Array. This may take a long time...
Jan 22 22:42:09 ... sorting Suffix Array chunks and saving them to disk...
Jan 22 22:47:18 ... loading chunks from disk, packing SA...
Jan 22 22:47:42 ... finished generating suffix array
Jan 22 22:47:42 ... generating Suffix Array index
Jan 22 22:49:38 ... completed Suffix Array index
Jan 22 22:49:38 ..... processing annotations GTF
terminate called after throwing an instance of 'std::bad_alloc'

      what():  std::bad_alloc
    /bin/sh: line 1: 186783 Aborted                 STAR --runThreadN 10 --runMode genomeGenerate --genomeDir /gpfs/ysm/scratch60/polimanti/ag2646/99H50_new_annotation/z10starindex75/ --genomeFastaFiles /gpfs/ysm/scratch60/polimanti/ag2646/Lawsonreference/genome.fa --sjdbGTFfile /gpfs/ysm/scratch60/polimanti/ag2646/Lawsonreference/genes.gtf --sjdbOverhang 75

我用谷歌搜索，发现这些错误可能源于内存分配，因此我从集群中有足够空间的空间运行。

此类作业的内存使用量由作业 ID 给出：

47861791 Array Job ID: 47861791_0

Cluster: farnam User/Group: ag2646/nicoli State: FAILED (exit code 134)
Nodes: 1 Cores per node: 10 CPU Utilized: 00:36:34 CPU Efficiency: 45.14% of 01:21:00 core-walltime Job Wall-clock time: 00:08:06 Memory Utilized: 25.64 GB Memory Efficiency: 25.64% of 100.00 GB.

我浏览了互联网并试图找出解决方案。

  1. 我尝试将线程数从 10 减少到 1，以减少计算内存问题。
  2. 我尝试通过使用以下标志来分配特定的内存限制：
    `limitGenomeGenerateRAM` 
    
        48000000000
        (3) --genomeChrBinNbits 16
        Still the error is creeping in.
        First few lines of my GTF file is chr12   UMMS    gene    6160446 6177944 .       -       .       gene_id "LL0000000001"; gene_name "a1cf";
    
    chr12   UMMS    exon    6160446 6161260 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6163727 6163869 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6165086 6165222 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6165305 6165498 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6167117 6167396 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6168940 6169037 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6169982 6170146 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6170412 6170650 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";
    chr12   UMMS    exon    6170731 6170861 .       -       .       gene_id "LL0000000001"; gene_name "a1cf"; transcript_id "ENSDART00000152292";

基因组fasta文件的一些行如下：

chr1
gatcttaaacatttattccccctgcaaacattttcaatcattacattgtc
atttcccctccaaattaaatttagccagaggcgcacaacatacgacctct
aaaaaaggtgctgtaacatgtacctatatgcagcaccactatatgagagc
ggcatagcagtgtttagtcacttggttgctttgtttatattaacttgaaa
gtgtgttttagctattgagtttaaacaaagggagcggtttacattgaatt
aaaggcaactactgatgggttgtgtaatgtttcaaagagctgttgcagca
tgagtggaaaataaaaccgtattagtgctgcctggcccagtttggcacaa
aatggagcgattccattaagagaacgattcagcataagtggaacagcTAA
AGtttatgaaaatttttaatctggatgtagagaatctcataacacagaaa

我试图提供尽可能多的细节，任何帮助都会有所帮助。

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