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北海舰队

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[交流] 血泪求助:Trinity的normalize 已有2人参与

鄙人最近用Trinity软件normalize一个trim后的数据,奈何一直报错如下:
Error, pairs.K25.stats is empty.  Be sure to check your fastq reads and ensure that the read names are identical except for the /1 or /2 designation. at /diag/home/aubzyy/trinityrnaseq_r20140717/util/insilico_read_normalization.pl line 865.
这是咋回事啊,俺是新手,跪求大侠出手相助!
我的数据是Pair-end。。。
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wizardfan

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it says that the name of sequences in your fastq can not match
In fastq format, Line 1 begins with a '@' character and is followed by a sequence identifier and an optional description (like a FASTA title line).
e.g. @HWI-EAS209_0006_FC706VJ:5:58:5894:21141#ATCACG/1
another entry would be something like
@HWI-EAS209_0006_FC706VJ:5:58:5894:21141#ATCACG/2
3楼2015-01-16 08:09:27
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匿名

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2楼2015-01-15 10:29:15
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北海舰队

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3楼: Originally posted by wizardfan at 2015-01-16 08:09:27
it says that the name of sequences in your fastq can not match
In fastq format, Line 1 begins with a '@' character and is followed by a sequence identifier and an optional description (like a FASTA  ...

Thank you. But my sequence starts like this:
@SRR1559285.21 HWI-ST531R:238:C346VACXX:5:1101:1193:2038 length=101
CCTTCATTCATGTGCTCACGCCAGTATGCAGCATTTTTAACAGCACAGTCCCAGT
+SRR1559285.21 HWI-ST531R:238:C346VACXX:5:1101:1193:2038 length=101
@CCFFFFFGHHGHIIJJJ4FHIJJGIIJJIIJIJJJJJHIJJJIIJJJHHJJJJI
@SRR1559285.22 HWI-ST531R:238:C346VACXX:5:1101:1126:2077 length=101
CCTATTCATTTACAGCAGAAGTGCTATTTCTTATTCATGATGTACAAGTCGTTGTTCCGTGAAAATGAAACAGAGACGGG
CGTCGTAGGATCCTTAGACGG
+SRR1559285.22 HWI-ST531R:238:C346VACXX:5:1101:1126:2077 length=101
CCCFFFFFHHGHHFIJJJHIIIHIJJJIJIJJIHJGGIFHIIGGIIJJFGHGEGHGIGGDGGGJJIIJGHIIIHJJHHHF
FD?BDBBBDCCCDDDDC:@@B
@SRR1559285.23 HWI-ST531R:238:C346VACXX:5:1101:1228:2083 length=101
CAGACGAATTATAGAAAGGCTGTTTAATTAGAAATTTACTTAATCCAAGTTTTGGCTGAAATGGATTGAAGTTTGTCCTG
GAATGTTTCGAACACATCTGT
+SRR1559285.23 HWI-ST531R:238:C346VACXX:5:1101:1228:2083 length=101
CCCFFFFFHFHGFIBHIJIJJJJGIGAHIEIHIHHIHIFIIIIIJJJEHHGIJIIHIIGJGIII@FHJGIGCAHEEHHFF
FFFCEEEECDDDBDDDDCD:@
@SRR1559285.24 HWI-ST531R:238:C346VACXX:5:1101:1246:2099 length=101
TGAAACTGAGGAATGTATGCCTCTCACGGTGACCCTGTCTCACCGTCTGAACCAAAAAATTGCTGAGCTTCGTAGAGATG
GTACATTCTGGTGGGCAAGAC
+SRR1559285.24 HWI-ST531R:238:C346VACXX:5:1101:1246:2099 length=101
@@@DF;DDBFFFFGGGHIJJAHGHIGEHGACGIJFIGIIGGIJJJHGIJIIJJEIIIIGGCACEEEFDEFFFDEC;@CCC
D@DCDFDDEEDCDDDB?@B<9
@SRR1559285.25 HWI-ST531R:238:C346VACXX:5:1101:1179:2100 length=101
Is this okay? I am a first year graduate, thank you for your patience.
4楼2015-01-19 13:40:24
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wizardfan

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小木虫: 金币+0.5, 给个红包,谢谢回帖
how sure you are that this is a pair-end data? Normally two files for pair-end data, named like ***_1.fastaq and ***_2.fastq. In ***_1.fastq, the headers of sequences are like ********/1, so as in the second file.
There is a public server for you to use Trinity https://galaxy.cbio.mskcc.org/
5楼2015-01-20 10:18:19
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