Q&A Apr 8, 2026

Protocol of yeast one hybrid assay using pLacZi2u/pJG4-5 systemVectorspLacZi2μ (AmpR in E.coli; Ura- in yeast) and pJG4-5 (or named pB42AD; AmpR in E.coli; Trp- in yeast) plasmids. 2μ origin of replication was PCR amplified from pEG202 and inserted into the HindIII and EcoRI sites of the reporter plasmid pLacZi (Clontech), resulting in pLacZi2μ vector.The coding sequence of DNA binding protein should be inserted into the MCS of pJG4-5 to generate the AD-fusion construct and the cis-element to be...

Q&A Apr 6, 2026

Cauda sperm collection protocolDissect out cauda epididymis from an adult mouse and transfer to a 35mm dish containing 1ml of pre-warmed Whitten’s Media (100 mM NaCl, 4.7 mM KCl, 1.2 mM KH2PO4, 1.2 mM MgSO4, 5.5 mM Glucose, 1 mM Pyruvic acid, 4.8 mM Lactic acid (hemicalcium), and HEPES 20 mM). Gently squeeze the epididymis to allow the caudal fluid to ooze out in the dish.  Incubate at 37°C for 15 minutes After incubation transfer the sperm containing media to a fresh tube and incubate for anoth...

Q&A Mar 31, 2026

Surgical Denervation of Interscapular Brown Adipose Tissue (iBAT) MaterialsAnesthetic agentSyringeInjection needleSterilized surgical instrument setsStraight forcepsCurved forcepsScissorsNote: To prevent contamination, it is recommended to use separate instrument sets for skin incision and for deeper tissue dissection. Instruments should be sterilized before use in different animals.Sutures: Pronova (blue, 4-0, 90 cm, taper-cut needle, 26 mm, strong curvature; 3975H)5% Hibitane (chlorhexidine; d...

Q&A Mar 30, 2026

Rat Lung scRNAseq DissociationLast updated 2026-03-30 by MSBRRaredon Lab Animals: SD Female and Male ~220/250grams Anesthetic: KX 75mg/5mg per kg body weight Pre-Op: Intraperitoneal heparin 0.15ml at 1000U/ml (critical to allow blood clearance from lung tissue) Setting: Biosafety hood for sterile cell isolation. Benchtop OK for non-sterile. Surgery:Abdominal incisionDiaphragm incision and tensionChest clamshellThymic dissection to expose great vesselsNeck dissection to expose tracheaCannulate tr...

Q&A Mar 23, 2026

Protocol: Investigating the Mechanism of MFN2 in Regulating Mitochondria-Associated Membranes (MAMs) and Cardiolipin in Vascular Dementia AbstractVascular dementia (VaD) is a progressive cognitive impairment caused by cerebrovascular disease, with mitochondrial dysfunction playing a pivotal role in its pathogenesis. Mitofusin 2 (MFN2) is a key protein mediating the tethering between the endoplasmic reticulum (ER) and mitochondria at specific contact sites known as mitochondria-associated membran...

Q&A Mar 18, 2026

Soil Extraction Plating for the Detection of Aspergillus terreus from Environmental SamplesAuthors and AffiliationsJan Schobert¹, Paul Illmer², Roya Vahedi-Shahandashti¹*, Cornelia Lass-Flörl¹*¹ ISHAM Working Group of Aspergillus terreus, ECMM Excellence Center of Mycology, Institute of Hygiene and Medical Microbiology, Medical University of Innsbruck, Austria² Institute of Microbiology, University of Innsbruck, AustriaAbstractThis protocol describes a selective soil extraction plating method fo...

Q&A Mar 17, 2026

Isolation of Extracellular Vesicles from Trichomonas vaginalisAbstractThis protocol describes the isolation of extracellular vesicles (EVs) from Trichomonas vaginalis using differential ultracentrifugation. Parasites are first cultured in TYM medium supplemented with heat‑inactivated horse serum until reaching the logarithmic growth phase. To induce EV release under serum‑free conditions, cells are washed and incubated for 4 hours in serum‑free TYM base. The conditioned medium is then clarified ...

Q&A Mar 16, 2026

2-Deoxy-D-glucose (2-DG) Treatment of Zebrafish EmbryosActivation of the Mondo Pathway via MicroinjectionDickmeis Laboratory IBCS-BIP/KIT, Karlsruhe, GermanyTechniqueZebrafish Embryo MicroinjectionApplicationActivation of MondoA/MLX pathway via 2-deoxy-D-glucoseModel OrganismDanio rerio (zebrafish)Document Version1.0Last Updated13/03/2026 1. Introduction and PrincipleThis protocol describes the use of the glucose analog 2-deoxy-D-glucose (2-DG) to selectively activate the Mondo pathway in zebraf...

Q&A Mar 6, 2026

 ATAC_qPCR Dissect embryos in Ringers solution. Single neural folds (˜3000 cells) contains enough cells for testing a few primer pairs.Transfer dissected samples to a microcentifuge tube (low binding).Rinse with 300uL Accumax to remove excess salt from RingersAdd 300uL of Accumax and incubate the tissue at RT for 20-30’, rocking at orbital shaker. During this period, gently pipette the tissue up and down to help tissue start dissociate.Centrifuge 3 min 600rcf at room temperature and withdraw liq...

Q&A Mar 2, 2026

Section I. Hazardous Reagent Safety & Preparation of key solutions (section is for both protocols) Mandatory Safety Rules (read primary MSDS and review EH&S guidelines before proceeding)Fume Hood Requirement: All weighing and mixing of dry powders (PFA and NaN3) must be performed inside a certified chemical fume hood.Respiratory Protection: Per SDS guidelines, wear a well-fitted N95 mask when weighing powders to prevent inhalation of toxic dust (NaN3 can be fatal if inhaled; PFA is a car...