转录组合并数据分析报告-454

上海众信生物技术有限公司

数据分析报告

2013年5月1日

XX转录组合并数据分析报告

目 录

1．分析要求 ....................................................................................................................................................3 2．分析流程及结果.......................................................................................................................................3 2.1 实验流程 ...............................................................................................................................................3 2.2 数据分析流程 ......................................................................................................................................4 2.3 原始数据 ...............................................................................................................................................4 2.4 序列拼接 ...............................................................................................................................................5 2.5 SSR分析................................................................................................................................................6 2.6 SNP分析 ...............................................................................................................................................6 2.7 BLAST注释.............................................................................................................................................7 2.8 GO分类 .................................................................................................................................................7 2.9 差异基因分析 ......................................................................................................................................9 2.10 UNIGENE比对 ....................................................................................................................................9 3．结果文件及说明.....................................................................................................................................10 3.1 软件参数设置 ....................................................................................................................................10 3.2 结果文件目录说明 ...........................................................................................................................11 4. 参考文献 ...................................................................................................................................................12 5. 附录（实验部分PROTOCOL） .......................................................................................................12 5.1 RNA提取 ............................................................................................................................................12 5.1.1 RNA precipitation .......................................................................................................................12 5.1.2 RNA wash.....................................................................................................................................13 5.1.3 RNA resuspension .......................................................................................................................13

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5.2反转录成CDNA ................................................................................................................................13 5.2.1 First-strand cDNA synthesis, set up 2 tubes for 2ug polyA RNA, for each tube: ............14 5.2.2 Second strand synthesis, for each tube:..................................................................................14 5.2.3 Phenol chloroform (Corrosive) isoamyl alcohol (PCI) and EtOH precipitation ............15 5.2.4 Gel separation.............................................................................................................................16 5.2.5 Gel extraction..............................................................................................................................16 5.2.6 Sonication ....................................................................................................................................17 5.3 454 LIBRARY PREPARATION ................................................................................................................17 5.3.1 Fragmentation of RNA...............................................................................................................17 5.3.2 Double-stranded cDNA Synthesis............................................................................................18 5.3.3 Fragment End Repair ................................................................................................................19 5.3.4 AMPure Bead Preparation........................................................................................................20 5.3.5 Adaptor Ligation.........................................................................................................................20 5.3.6 Small Fragment Removal ..........................................................................................................21 5.3.7 Library Quantitation ..................................................................................................................21 5.3.8 cDNA Library Quality Assessment ..........................................................................................23 5.3.9 Preparing Working Aliquots .....................................................................................................23

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XX转录组合并数据分析报告

1．分析要求

对经预处理的四个组织的454测序结果进行合并拼接、以及对此拼接结果进行注释、SNP分析、SSR分析、GO功能分类等分析。

2．分析流程及结果

2.1 实验流程

RNA提取 反转录成cDNA 454测序 454 Library制备

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2.2 数据分析流程

2.3 原始数据

实验共得到4个组织各1/2 run数据，合计2个run的测序数据。数据预处理已在上次报告中给出，这里对经过预处理后合并的数据进行reads长度频数统计，详细结果见reads_statistics.xls中original_reads_stat sheet。图2.3.1为reads长度分布示意图，表2.3.1给出预处理后reads基本统计信息。

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