Bulk RNAseq of chronic LPS treated female mouse pituitary across doses
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http://datadryad.org/dataset/doi%253A10.5061%252Fdryad.x69p8czrd
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To investigate the mechanisms of chronic inflammation on gonadotropin secretion we performed bulk RNA-seq on pituitaries from female mice chronically exposed to lipopolysaccharide for 6 weeks.
Methods
Pituitaries were dissected between 10 to 11 am from mice after 6 weeks of twice weekly i.p. injections and when they reache diestrous. The pituitaries were frozen in RNAlater (Qiagen) at minus 80 degrees Celsius.
RNA was harvested using Rneasy mini plus kit (Qiagen). 1.3 ug of total RNA was used for the construction of sequencing libraries.
RNA libraries for RNA-seq were prepared using SMARTER mRNA-Seq Library Prep Kit following manufacturer's protocols.
RNA-seq
CLC Genomics Workbench v 11.0.1
Sequence reads were trimmed for adaptor sequence/low-quality sequence using CLC genomic benchwork (parameter- Quality limit: 0.05)
Trimmed sequence reads were mapped to GrCM38/mm10 using CLC genomic benchwork (parameters- mismath cost: 2 insertion cost: 3 deletion cost: 3 length fraction: 0.8 similarity fraction: 0.8)
Read count extraction and normalization were performed using CLC genomic benchwork
创建时间:
2024-01-24



