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| Status |
Public on Oct 28, 2016 |
| Title |
Extensive cryptic splicing upon loss of RBM17 and TDP43 in neurodegeneration models |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Translating ribosome affinity purification technology was used to isolate mRNAs from cerebellar Purkinje neurons from control (Pcp2-BacTrap; Rbm17 f/+) and mutant (Pcp2-BacTRAP; Pcp2-Cre; Rbm17 f/-) mice.
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| Overall design |
RNA isolation was performed when animals were four-weeks-old (n=3 animals per genotype). Using NuGEN Ovation RNA-Seq System v2, purified double-stranded cDNA was generated from 10 ng of total RNA and amplified using both 3’ poly (A) selection and random priming. 2 µg of each sample was sheared using the Covaris S2 focused-ultrasonicator following the manufacturer’s protocol to obtain a final library with insert size of 400 bp. The sheared samples were quantified using the NanoDrop ND-1000 spectrophotometer and Invitrogen Qubit 2.0 DNA quantitation assay. The fragment sizes were confirmed on the Agilent Bioanalyzer to verify proper shearing. A double-stranded DNA library was produced using Illumina TruSeq DNA library preparation system and the sequencing was run on a HiSeq 2500 system.
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| Contributor(s) |
Zoghbi HY, Liu Z |
| Citation(s) |
28007900 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R37 NS027699 |
MOLECULAR STUDIES OF SPINOCEREBELLAR ATAXIS TYPE I |
BAYLOR COLLEGE OF MEDICINE |
HUDA Y ZOGHBI |
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| Submission date |
Mar 08, 2016 |
| Last update date |
May 15, 2019 |
| Contact name |
Zhandong Liu |
| E-mail(s) |
zhandonl@bcm.edu
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| Phone |
8328248878
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| Organization name |
Baylor College of Medicine
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| Street address |
1250 Moursund St., STE NR-1325
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| City |
Houston |
| State/province |
TEXAS |
| ZIP/Postal code |
77030 |
| Country |
USA |
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| Platforms (1) |
| GPL17021 |
Illumina HiSeq 2500 (Mus musculus) |
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| Samples (6)
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| Relations |
| SRA |
SRP071321 |
| BioProject |
PRJNA314719 |
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