SAGE Library SM127

Classification

Library Name SM127
Species Mus musculus
Strain C57BL/6J
Mutation Wild Type
Location Lung - Epithelium
Sex Unknown
Age E11.5
Developmental Stage Theiler Stage 19
Organsim Condition

Sample Collection

Tissue Dissection Photos

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With the aid of a dissecting microscope, whole lungs were manually removed from embryos submerged in phosphate buffered saline (PBS). To dissociate the epithelium, lungs were subsequently placed in 500 uL of 1% trypsin in Hanks solution. After 90 minutes of incubation at 4ÂșC, the digestion was stopped with the addition of 1 mL of 20% fetal bovine serum (FBS) in Hanks solution plus a small amount of DNase I powder. Tungsten needles were used to tease away the epithelium which was collected in tubes containing Trizol. Tissue from several litters was collected to obtain enough RNA for SAGE library construction. No more than two litters were dissected on the same day, with each litter taking about 1hour to manually dissocitate after trypsin digestion.

Creative Commons License
This work is licensed under a Creative Commons Attribution 2.5 License.
Lab BCCRC: Pamela Hoodless Lab
Number of Animals 64

Library Construction

SAGE Protocol SAGE (LongSAGE 21 bp)
Starting RNA 2.5 ug

Tag Extraction

Total Raw Tags (no quality cut-off) 112,307
Total Raw Tags at Q95 88,677
Total Raw Tags at Q99 84,605
Unique Tags 39,069

Data

Library Metadata Library Metadata XML
Tag Counts at Quality 0.95 cut-off tagcounts_SM127_Q95.txt
Tag Counts at Quality 0.99 cut-off tagcounts_SM127_Q99.txt
Clustered Tag Counts and P-Value tagclustered_SM127_Q99.txt

Data Analysis

DiscoverySpace

Data is available for analysis using DiscoverySpace, a cross-platform application that allows comparison between SAGE libraries, tag to gene mapping and results from public datasources.

SAGE Library Help

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