miRNA is extracted using Ambion’s nirvana miRNA isolation kit. This kit uses a modified fibreglass membrane filter method for a quick recovery of total RNA including miRNA from samples. In brief, the sample is first lysed in a denaturing lysis buffer and treated with acid-phenol:chloroform, then total RNA is purified over a glass-fiber filter. miRNA is further isolated from total RNA via gel purification. The quality of RNA is confirmed by quantifying the amount of RNA, so as to ensure the reliability of downstream experiments. For FFPE samples, Ambion’s RecoverAll Total Nucleic Acid Isolation Kit is used to extract miRNA. This kit is specifically designed to isolate DNA, RNA from FFPE samples, which also includes all the miRNA. Eluted miRNA can be directly used for miRNA microarray. In the case of anticoagulant whole blood sample, Ambion’s RiboPure-Blood Kit is used to extract miRNA. In addition, Nirvana PARIS kit can be used to extract natural protein and total RNA including all the small non-coding RNAs (miRNA, nRNA, snoRNA). This allows researchers be able to examine the correlation between miRNA expression level and protein level.
Sample Requirement
– Fresh blood/tissue/cultured cells (collected within 0.5-2 hours without cryopreservation)
– Tissue/isolated white blood cells/cultured cells preserved in liquid nitrogen
– Trizol properly treated tissue/isolated white blood cells/cultured cells (Cryopreservation within 3 days)
– Formalin fixed paraffin embedded blocks/sections.
Service Workflow
1. miRNA isolation
2. miRNA quality check
3. Experimental report
Deliverables
1. Experiment record
2. Quality check report
