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yuehedou

木虫 (小有名气)

[求助] miRNA高通量测序分析时拷贝数的选取问题

分析高通量测序的miRNA数据时,通过质控的序列在进入后续分析(conserved,novel等的分析预测)时,对拷贝数(count数)有没有要求?
我在文献中看到有人全部拿去分析,有人去除了单拷贝,还有人甚至只用10拷贝以上的序列。
欢迎指教、讨论!
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growlywolf

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【答案】应助回帖

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yuehedou: 金币+20, ★★★很有帮助, 谢谢! 2014-04-27 14:53:12
我一般取5个reads以上的
或者取TPM>1的
2楼2014-04-26 20:19:15
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XOooZzz

银虫 (小有名气)

miRBase有这样的建议:
(1) Multiple reads (10–20 are commonly used cutoffs) support the presence of the mature microRNA (preferably from multiple independent experiments);
(2) The reads map to an extended sequence region (e.g. an assembled contig), and the sequence
flanking the putative mature microRNA folds to form a microRNA precursor-like hairpin with strong pairing between the mature microRNA and the opposite arm. Reads that map very many times
to a genome sequence should be discarded;
(3) Mapped reads do not overlap other annotated transcripts (i.e. there is no evidence that the short readsmay represent fragments of mRNAs or other known RNA types);
(4) Reads mapping to a locus support consistent processing of the 5'-end of the mature sequence (for example, the majority of reads overlapping a given mature microRNA annotation should have the same 5'-end; the 3'-end may be significantly more variable); and
(5) Ideally, reads will support the presence of mature sequences from both arms of the predicted hairpin (so-called miR and miR* sequences), and the putative mature sequences should base-pair with the correct 3'-overhang.

miRBase: integrating microRNA annotation and deep-sequencing data
3楼2014-04-27 10:47:13
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yuehedou

木虫 (小有名气)

引用回帖:
2楼: Originally posted by growlywolf at 2014-04-26 20:19:15
我一般取5个reads以上的
或者取TPM>1的

谢谢!
每天都为自己的无知而羞耻!
4楼2014-04-27 14:52:49
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yuehedou

木虫 (小有名气)

引用回帖:
3楼: Originally posted by XOooZzz at 2014-04-27 10:47:13
miRBase有这样的建议:
(1) Multiple reads (10–20 are commonly used cutoffs) support the presence of the mature microRNA (preferably from multiple independent experiments);
(2) The reads map to an e ...

很详细,谢谢!
每天都为自己的无知而羞耻!
5楼2014-04-27 14:54:29
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