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| Status |
Public on Feb 10, 2023 |
| Title |
Characterizing role of striated-muscle specific ribosomal protein L3-like (Rpl3L) in mouse hearts |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Overall goal: To elucidate the function served by Rpl3L in heart. Purpose of analysis: To determine effects of Rpl3L-deficiency on translational mRNA profile in heart. Experimental structure: Polysomes and monosomes were fractionated from cardiac ventricles and associated RNA extracted for library generation and differential gene expression analyses between wild-type and Rpl3L-null samples, using standard RNAseq pipeline.
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| Overall design |
Whole ventricles from wild-type control and Rlp3L-null experimental hearts were pulverized and subsequently manually homogenized using Dounce homogenizers on ice in polysome lysis buffer (10 mM Tris-HCl at pH 7.5, 100 mM KCl, 5mM MgCl2, 0.5% w/ve sodium deoxycholate, 1% v/v IGEPAL CA-630, 10 mM DTT, 150 µg/ml cycloheximide, 1U/µl SUPERase In RNase inhibitor, and 1X HALT protease inhibitor in UltraPure molecular grade H2O). After serial centrifugation, supernatant from final 5-min spin at 16,000 x g at 4°C was layered on top of a 15 to 60% sucrose gradient column and centrifuged at 38,000 RPM for 125 min in a Optima XPN-90 ultracentrifuge (Beckman Coulter + SW 60 Ti swing bucket rotor). Total RNA from monosomal+polysomal fractions was extracted and subjected to downstream sequencing and bioinformatics analyses.
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| Contributor(s) |
Grimes K, Prasad V, Molkentin JD |
| Citation(s) |
36733907 |
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| Submission date |
Oct 16, 2022 |
| Last update date |
Feb 11, 2023 |
| Contact name |
Jeffery D Molkentin |
| E-mail(s) |
vik.jd.geosubmissions@gmail.com
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| Organization name |
Cincinnati Children's Hospital Medical Center
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| Department |
Molecular Cardiovascular Biology
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| Lab |
Molkentin Lab
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| Street address |
240 Albert Sabin Way MLC 7020
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| City |
Cincinnati |
| State/province |
Ohio |
| ZIP/Postal code |
45229 |
| Country |
USA |
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| Platforms (1) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA891061 |
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