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RNA Extraction from Ears and Draining Lymph Nodes of Mice Infected with Leishmania amazonensis
- Source :
- Bio-protocol, Bio-protocol, Bio-protocol LCC, 2020, 10 (11), ⟨10.21769/BioProtoc.3633⟩, Bio Protoc, Bio-protocol, 2020, 10 (11), ⟨10.21769/BioProtoc.3633⟩
- Publication Year :
- 2020
- Publisher :
- HAL CCSD, 2020.
-
Abstract
- International audience; Parasites of the genus Leishmania infect the mammalian hosts, including mice and humans and cause cutaneous or visceral leishm aniasis depending upon the parasite species transmitted by the vector sandfly. Leishmania amazonensis is one of the Leishmania species responsible for th e cutaneous form of the disease. We have inoculated with these parasites the ear dermis of mice. RNA preparations were performed from fragmented tissues using a buffer containing guanidin isothiocynate (RLT buffer, RNeasy Mini Kit, Qiagen, SAS, France) and β mercaptoethanol. Both reagents facilitate the isolation of intact RNA from tissues and the use of the RNeasy Kits present with several advantages that facilitate the isolation of pure non degraded total RNA: i) T his method allows to avoid the presence o f phenol in the RNA extraction buffer, commonly used in alternative protocols; ii) Moreover Diethylpyrocarbonate (DEPC) treatment of glassware, to avoid RNAses contamination of the samples, is not required with this protocol; iii) F inally, it is a fast pro cedure and the isolated total RNA may be concentrated in a small volume thus facilitating its use for downstream experimental procedures.
- Subjects :
- Mouse
Strategy and Management
[SDV.BC.BC]Life Sciences [q-bio]/Cellular Biology/Subcellular Processes [q-bio.SC]
Industrial and Manufacturing Engineering
Microbiology
Dermis
[SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases
medicine
Methods Article
Parasite hosting
Vector (molecular biology)
Ear pinna dermis
Lymph nodes
biology
Mechanical Engineering
Metals and Alloys
RNA
[SDV.MHEP.EM]Life Sciences [q-bio]/Human health and pathology/Endocrinology and metabolism
biology.organism_classification
medicine.disease
3. Good health
Sandfly
RNA isolation
[SDV.GEN.GA]Life Sciences [q-bio]/Genetics/Animal genetics
medicine.anatomical_structure
Visceral leishmaniasis
[SDV.GEN.GH]Life Sciences [q-bio]/Genetics/Human genetics
RNA extraction
Lymph
Leishmania amazonensis
Subjects
Details
- Language :
- English
- ISSN :
- 23318325
- Database :
- OpenAIRE
- Journal :
- Bio-protocol, Bio-protocol, Bio-protocol LCC, 2020, 10 (11), ⟨10.21769/BioProtoc.3633⟩, Bio Protoc, Bio-protocol, 2020, 10 (11), ⟨10.21769/BioProtoc.3633⟩
- Accession number :
- edsair.doi.dedup.....cb543a008728542939a54e963f0bc92d