Individual E9 RV+OFT pieces were dissected manually as described above and stored in QIAzol lysis reagent (Qiagen) at −80°C until RNA isolation using miRNeasy mini kit (Qiagen). Individual E9 second heart field-enriched torso pieces without heart were dissected manually and stored at −80°C until RNA isolation using miRNA purification kit from Norgen Biotek. After quantification, samples were shipped overnight on dry ice to LC Sciences (Houston Texas) who assayed the samples miR-17 and miR-20a (using Taqman miRNA assays Ids 2308, 0580) and control RNAs snoRNA202 and noRNA234 (assay Ids 1232, 1234). Three stage-matched pairs of controls and mutants were used for each genotype assayed; each of these assayed in triplicate for each assay, along with a no template control.
Mirna purification kit
Manufactured by Norgen Biotek
The MiRNA purification kit is a laboratory tool designed to isolate and purify microRNA (miRNA) from various biological samples. The kit utilizes a specialized extraction and purification protocol to efficiently capture and concentrate miRNA molecules for further analysis and downstream applications.
Automatically generated - may contain errors
2 protocols using mirna purification kit
1
Cardiac Development microRNA Profiling
2
Extraction and Purification of RNA from Solid Tumors and Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
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From MSKCC's prospectively maintained tissue bank, fresh frozen tumors or formalin fixed, paraffin embedded (FFPE) tissues mounted on glass slides were obtained for each patient. Solid tumor samples were flash frozen, preserved in RNAlater (Thermofisher AM7024) to minimize RNA degradation, cut and measured to roughly the same size, and grounded with disposable pestles (Sigma Z359947) before being subjected to RNA extraction using the Norgen miRNA purification kit (21300) per manufacturer's instructions. Extracted RNA was then bioanalyzed for integrity prior to library preparation. To extract RNA from FFPE tissue slides, a pathologist (J.S.) reviewed the H&E slides for visual confirmation of tumor cells, and areas containing tumor cells were encircled for extraction. Tumor cells were scraped from corresponding unstained slides with a surgical blade, and miRNAs were isolated as described above. For the serum cohort, RNA was extracted from 1mL of serum using Norgen Plasma/Serum Circulating and Exosomal RNA Purification Kit (Slurry Format) (42800).
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