{"Id":"ad498047-df9c-4a7e-b8a4-c56d1e981355","CanonicalId":"dx.doi.org/10.17504/protocols.io.bwebpban","Url":"https://www.protocols.io/view/a-systematic-comparison-of-protocols-for-recovery-bwebpban","Name":"A Systematic Comparison of Protocols for Recovery of High-Quality RNA from Human Islets Extracted by Laser Capture Microdissection ","Description":"The isolation of high-quality RNA from endocrine pancreas sections represents a considerable\nchallenge largely due to the high ribonuclease levels. Laser capture microdissection (LCM) of\nmammalian islets, in association with RNA extraction protocols, has emerged as a feasible approach\nto characterizing their genetic and proteomic profiles. However, a validated protocol to obtain highquality\nRNA from LCM-derived human pancreas specimens that is appropriate for next-generation\nsequencing analysis is still lacking. In this study, we applied four methods (Picopure extraction\nkit, Qiazol protocol, Qiazol + Clean-up kit, and RNeasy Microkit + Carrier) to extract RNA from\nhuman islets obtained from both non-diabetic individuals and patients with type 2 diabetes who had\nundergone partial pancreatectomy, as well as handpicked islets from both non-diabetic and diabetic\norgan donors. The yield and purity of total RNA were determined by 260/280 absorbance using\nNanodrop 100 and the RNA integrity number with a bioanalyzer. The results indicated that among\nthe four methods, the RNeasy MicroKit + Carrier (Qiagen) provides the highest yield and purity.\n","ResourceType":"Protocol","ParentResourceType":null,"DefiningManuscriptId":"","Publications":[],"Contributors":[{"Role":{"CanonicalId":"hirn-cv-ResourceRole-producer","Url":null,"Name":"Producer","OntologyId":null},"HirnUserId":"rohit_kulkarni@hirnetwork.org","FirstName":"Rohit","LastName":"Kulkarni","EmailAddress":"rohit.kulkarni@joslin.harvard.edu","Organization":"Joslin Diabetes Center","Consortia":["CBDS","CTAR"]}],"Vendor":{"id":null,"Name":"","Url":null},"VendorResourceName":null,"VendorResourceUrl":"","VendorCatNumber":"","VendorLotNumber":"","DateCreated":"2021-12-06T19:24:48.1094094+00:00","DateModified":"2022-07-01T03:36:55.5976354+00:00","VersionId":"1","ModelVersion":"4.0","ProtocolData":{"ProtocolType":{"CanonicalId":null,"Url":null,"Name":null,"OntologyId":null},"DocumentId":null,"ProtocolSpecies":[],"DocumentTopic":[]}}