User contributions for Admin

19:5219:52, 1 February 2021 diff hist +28‎ m Fab preparation ‎ →‎References current Tag: Visual edit
19:5219:52, 1 February 2021 diff hist +4,052‎ N Fab preparation ‎ Created page with "== Buffers Required: == <pre>Note: THe volume of buffers prepared is dependent on the starting volume of antibody-containing media. Adjust volumes accordingly. </pre> =====..."
19:5119:51, 1 February 2021 diff hist +6,845‎ N Immunoprecipitation ‎ Created page with "== Phosphoprotein Immunoprecipitation == Receptor activation by ligand addition or cross-linking, followed by immunoprecpitation of phosphoproteins, can be used it identify t..." current
19:5119:51, 1 February 2021 diff hist +11,652‎ N Western Blotting ‎ Created page with "This is a generic protocol for western blotting. Optimized procedures for specific blots are listed at the end of this protocol. = Cell Preparation and Lysis = #Pr..." Tag: Visual edit: Switched
19:4919:49, 1 February 2021 diff hist −18‎ m Main Page ‎ →‎DNA/Cloning Tag: Visual edit
19:4419:44, 1 February 2021 diff hist +2,419‎ N Transformation ‎ Created page with "=== Protocol === #Use either 10-50 ng of DNA in a volume between 10 and 25 μL or entire ligation reaction and place the tube on ice. #Thaw 100 μL of competent ''E...." current
19:4419:44, 1 February 2021 diff hist +1,499‎ N Generating Minicircles ‎ Created page with "= Protocol = *Grow an overnight culture of ZYCY10P3S2T containing the appropriate minicircle parental plasmid in 1-5 mL TB + Kan **Recipe for 50 mL Terrific Broth: ***1.2 g Ye..." current Tag: Visual edit: Switched
19:4319:43, 1 February 2021 diff hist −2‎ Neon® Transfection System ‎No edit summary current Tag: Visual edit
19:4319:43, 1 February 2021 diff hist 0‎ N File:Tip.png ‎No edit summary current
19:4319:43, 1 February 2021 diff hist 0‎ N File:Chamber.png ‎No edit summary current
19:4219:42, 1 February 2021 diff hist +8,092‎ N Neon® Transfection System ‎ Created page with "The following is a general protocol for using the Neon® Transfection System, along with specifics for optimized use with common cell lines found in the Heit Lab. =Introducti..."
19:4219:42, 1 February 2021 diff hist +2,419‎ N E. coli Transduction ‎ Created page with "=== Protocol === #Use either 10-50 ng of DNA in a volume between 10 and 25 μL or entire ligation reaction and place the tube on ice. #Thaw 100 μL of competent ''E...." current
19:4219:42, 1 February 2021 diff hist +3,222‎ N Competent e coli ‎ Created page with "= TFB Protocol for DH5a = #Inoculate a single colony from an LB plate into 5ml of LB medium in a plating tube OR inoculate about 1 µL of competent ''E coli ''i..." current
19:4119:41, 1 February 2021 diff hist +2,892‎ N Quick 'n' Easy Competent E. coli ‎ Created page with "This protocol can be used to generate competent E. coli of the following strains: *DH5a *DH1 *JM109 This will '''NOT''' work for BL-21 nor ZYCY10P3S2T (minicircle) strains =..." current
19:4119:41, 1 February 2021 diff hist +3,053‎ N Gibson Assembly ‎ Created page with "Gibson Assembly, also known as enzymatic or chew-back assembly, is a powerful tool used for DNA cloning. It allows you to manipulate almost any fragment of DNA in any location..."
19:4119:41, 1 February 2021 diff hist 0‎ N File:Ga.png ‎No edit summary current
19:4019:40, 1 February 2021 diff hist +1,059‎ N Ligation ‎ Created page with "=== Protocol === #Determine the amount of plasmid and insert to be used through the equation X ng plasmid/[(1 M) X bp] = X ng insert/[(X M) X bp]. It is best to have a pasmid..." current
19:4019:40, 1 February 2021 diff hist +2,001‎ N Digests ‎ Created page with "=== Protocol === Add the following reagents into a round-lidded PCR tubes: {| cellspacing="1" cellpadding="1" border="1" style="width: 506px; height: 142px;" |- ! scope=..."
19:4019:40, 1 February 2021 diff hist +1,198‎ N PCR ‎ Created page with "=== Protocol for Phusion === *Add the following volumes to a round-lidded PCR tube: {| cellspacing="1" cellpadding="1" border="1" style="width: 326px; height: 225px..." current
19:4019:40, 1 February 2021 diff hist +726‎ N Colony PCR ‎ Created page with "= Protocol = #Mark a clean agar plate (with required antibiotic) with a grid; make sure top is clearly defined. #Prepare PCR tubes containing 10μl PCR reaction mix/tube. La..." current
19:3919:39, 1 February 2021 diff hist +5,723‎ N Gentamicin Protection Assay ‎ Created page with "== Introduction == The gentamycin protection assay is used to monitor the killing or growth of pathogenic bacteria by phagocytes such as macrophages and neutrophils. This assa..." current
19:3919:39, 1 February 2021 diff hist +4,912‎ N Inside-out Labelling of Bacteria ‎ Created page with "Many phagocytosis assays require that bacteria be labelled such that internalised bacteria can be clearly delineated from those merely bound to the cell surface. This is achie..." current
19:3919:39, 1 February 2021 diff hist +2,221‎ N Labelled E coli ‎ Created page with "This method will produce highly-fluorescent, heat-killed ''E. coli'' for phagocytosis assays. This assay has been optimised for ML35, but should work for other strains of ''E...." current
19:3919:39, 1 February 2021 diff hist +3,829‎ N Primary Human Macrophages ‎ Created page with "= Isolation & Culturing Primary Human Macrophages = Human macrophages can be prepared from anticoagulant-treated peripheral blood mononuclear cells. By culturing mo..." current
19:3819:38, 1 February 2021 diff hist +3,801‎ N Phagosome Isolation ‎ Created page with "This isolation is based on the protocol of Lonnbro ''et al<ref name="Pubmed Central">http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2453110&amp;amp;amp;tool..." current
19:3819:38, 1 February 2021 diff hist +925‎ N Synchronised Phagocytosis ‎ Created page with "Synchronised phagocytosis is used to ensure that all observed phagocytosis events in a maple have occurred after a similar amount of time. This is useful for microscopy..." current
19:3819:38, 1 February 2021 diff hist +4,272‎ N Preparation of Silica-Magnetic Beads ‎ Created page with "Silica (SiO2) beads and silica-coated magnetic beads are a substrate for the attachments of a broad range of biomolecules through polar interactions. Lipids,..." current
19:3819:38, 1 February 2021 diff hist +3,108‎ N Opsonization ‎ Created page with "Opsonization allows phagocytes to efficiently recognize particles, by coating them with opsonins such as IgG isotype antibodies. This page contains the protocols for opsonisin..." current
19:3719:37, 1 February 2021 diff hist +691‎ N Water Bath Antibiotic Solution ‎ Created page with "= Reagents Needed = *Copper Sulfate (Cu2SO4) *EDTA *Dish soap or other detergent (i.e. Tween/Triton) = Preparation = Fill water bath with tap water (typ..." current
19:3719:37, 1 February 2021 diff hist +1,860‎ N Agarose Gels ‎ Created page with "==== Protocol ==== ===== For a 0.7% gel: ===== #Set up gel-casting dock. #Add 0.14 g of agarose to a 250mL Erlenmeyer flask. #Add 400 µL of 50X TAE and 20 ml of d..."
19:3719:37, 1 February 2021 diff hist +675‎ N Antibiotic Plates ‎ Created page with "=== Protocol === #Add 20 g of LB agar to a 1 L flash. #Fill flask to the 500 mL line with dH2O. #Autoclave the flask. Note: have a water bath set to 50°C. #Whe..." current
19:3719:37, 1 February 2021 diff hist +988‎ N Antibiotics ‎ Created page with "== Protocol == ==== Ampicillin ==== Preparation of 5 mL of 100 mg/mL: #Add 0.5 g of ampicilin into a 12 mL tube. #Add 2.5 mL of ddH2O into the tube and mix g..."
19:3619:36, 1 February 2021 diff hist +2,253‎ N J774 Cell Transfection ‎ Created page with "= Protocol = ===== Cell Culture ===== #Prepare a 6-well plate by adding 2 ml of J774 Cell media per well. If coverslips are required, add one coverslip/well prior to add..."
19:3619:36, 1 February 2021 diff hist +2,285‎ N Raw Cell Transfection ‎ Created page with "= Protocol = ===== Cell Culture ===== #Prepare a 6-well plate by adding 2 ml of Raw Cell media per well. If coverslips are required, add one coverslip/well prior to addi..." current
19:2619:26, 1 February 2021 diff hist +1,284‎ N Freezing and Thawing Cells ‎ Created page with "= Freezing Mammalian Cells = #In a T75 flask, grow cells to 80-90% confluency, or 75-80% density for suspended cells. #Collect cells (trypsinize/scrape adherent cells, etc)...." current
19:2519:25, 1 February 2021 diff hist +2,796‎ N Primary Macrophage Culture ‎ Created page with "= Isolation and Culture of Mouse Bone Marrow-Derived Macrophages = === Protocol === ===== Materials ===== *Dulbecco's minimal essential medium (DMEM) from Invitroge..."
19:2519:25, 1 February 2021 diff hist +1,380‎ N Bacterial Growth Media ‎ Created page with "=== LB Broth === To 200 mL ddH2O add: *Pre-made: **6.25g Miller LB powder *'''''OR''''' **2.5g Tryptone **2.5g NaCl **1.25g Yeast extract..." current
19:2519:25, 1 February 2021 diff hist +2,226‎ N Cell Culture Guidelines ‎ Created page with "= Maintenance of Common Cell Lines = {| width="95%" border="1" cellpadding="1" cellspacing="1" |- | '''Cell Type''' | '''Media''' | '''Maintenance''' | '''Splitting''' | '''N..." current
19:2519:25, 1 February 2021 diff hist +4,126‎ N Common buffers ‎ Created page with "= Cell Culture = ===== 1x PBS ===== To 800ml ddH2O add: *8g NaCl *0.2g KCl *1.44g Na2HPO4 (sodium phosphate dibasic) ..." current Tag: Visual edit: Switched
19:2419:24, 1 February 2021 diff hist −62‎ m Main Page ‎ →‎Lab Operations Tag: Visual edit
19:2319:23, 1 February 2021 diff hist +3,139‎ N Setting Up Network Drives ‎ Created page with "The laboratory has a server on which all work-related files must be stored. Before you can access these folders you must get a user ID and password from Dr. Heit. Please..." current
19:2319:23, 1 February 2021 diff hist +2,828‎ N Saving Experimental Files ‎ Created page with "= Logging Computer Files = It is not uncommon for experiments to generate large quantities of computer files containing raw data, analyzed data, statistics, intermediary stag..." current
19:2319:23, 1 February 2021 diff hist +7,003‎ N Labbook Guidelines ‎ Created page with "= Preamble = Your laboratory notebook is an important tool that goes well beyond research management and can have important implications for issues ranging from intellect..." current
19:2219:22, 1 February 2021 diff hist +1,668‎ N Exit Protocol ‎ Created page with "It is critical that members leaving the lab properly archive all materials so that they can be found and used by others. This is critical to the success of both yourself..."
19:2219:22, 1 February 2021 diff hist +6,837‎ N Entrance Protocol ‎ Created page with "Beginning in a new lab can be an overwhelming experience. This guide is intended to aid new members in starting up as quickly as possible. = Accounts & Certificatio..."
19:2119:21, 1 February 2021 diff hist +5,494‎ N Reducing Photobleaching ‎ Created page with "Photobleaching is a common issue with both fixed and live samples. Proper sample preparation, and the addition of photoprotectants, can dramatically reduce the degree of photo..." current Tag: Visual edit: Switched
19:1819:18, 1 February 2021 diff hist +673‎ N Create new ‎ Created page with "= How to Create a New Page = #On the main page, in the index section, add the title of your new page. #Highlight the new title, then click the link icon on the..." current
19:1719:17, 1 February 2021 diff hist +2,581‎ N Editing ‎ Created page with "= Instructions for Creating or Editing Pages = To make the pages consistent, and protocols easy to follow, please follow these guidelines: #Structure the protocol in the ord..." current Tag: Visual edit: Switched
18:5618:56, 1 February 2021 diff hist +10,121‎ Main Page ‎No edit summary
18:4218:42, 1 February 2021 diff hist 0‎ N File:Logo.png ‎No edit summary current

User contributions for Admin

1 February 2021

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