We narrowed to 658 results for: otos
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TypeProtocol...Watch the protocol video below to learn how to inoculate bacteria in liquid culture. Protocol Prepare liquid...Protocol for Inoculating a Bacterial Culture Protocols... Protocols Inoculating a Liquid Bacterial Culture Inoculating...plasmid DNA for experimental use. The following protocol is for inoculating an overnight culture of liquid...cloudy haze in the media (see right). Notes: Some protocols require bacteria to be in the log phase of growth...growth. Check the instructions for your specific protocol and conduct an OD600 to measure the density of...
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Protocol - pLKO.1 – TRC Cloning Vector
TypeProtocol... Protocols pLKO.1 Protocol pLKO.1 - TRC Cloning Vector Addgene Plasmid 10878. Protocol Version ... RNAi Tools page. E.2 Protocol for Producing Lentiviral Particles This protocol is for transfection in...Addgene's protocol for using the pLKO.1 cloning vector, a backbone used by the RNAi consortium for targeting...Copyright Addgene 2006, All Rights Reserved. This protocol is provided for your convenience. See “warranty...Lentiviral Particles E.1 Recommended materials E.2 Protocol for producing lentiviral particles F. Infecting...Determining the optimal puromycin concentration F.3 Protocol for lentiviral infection and selection G. Safety...competent DH5 alpha cells, following manufacturer’s protocol. Plate on LB agar plates containing 100 μg/mL ... -
Protocol - How to Perform Sequence Analysis
TypeProtocol... Protocols Sequence Analysis Sequence Analysis of a Plasmid... -
Protocol - Over-Agar Antibiotic Plating
TypeProtocol...concentration. Last Update: Oct. 27, 2017 Protocol Video Watch the protocol video below to learn how to spread... Protocols Streaking Bacteria on LB Agar Plate Over-Agar ...Plasmid DNA from Bacterial Culture Introduction This protocol describes methodology for plating antibiotic over-agar...separate batches of antibiotic-containing agar. This protocol will focus specifically on selection with carbenicillin...transformation page for a detailed heatshock transformation protocol. Plate 50 µL of transformed E. coli /rescue media... -
Virus Protocol - Generating Stable Cell Lines
TypeProtocol... Virus Blog Posts Mol Bio Protocols Viral Service Introduction This protocol can be used to generate stable...Protocol to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector... Protocols Generating Stable Cell Lines with Lentivirus Generating Stable Cell Lines with Lentivirus...lentiviral vectors deliver antibiotic resistance. This protocol was established using 293T cells but can be adapted...FBS lot numbers to find one suitable with your protocols. FBS can be purchased already heat-inactivated... -
Protocol - How to Create a Bacterial Glycerol Stock
TypeProtocol...stored stably at -80°C for many years. Protocol Video Watch the protocol video below to learn how to create...Protocol for creating a glycerol stock, including additional tips for long-term storage. Protocols... Protocols Creating... -
Plasmid Modification by Annealed Oligo Cloning (with Protocols)
TypeProtocol... Protocols Plasmid Modification by Annealed Oligo Cloning...using a commercially available kit or standard protocol. Anneal oligos The oligos should be resuspended... -
Plasmid Cloning by Restriction Enzyme Digest (with Protocols)
TypeProtocol... Protocols Subcloning Plasmid Cloning by Restriction Enzyme... -
Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps
TypeProtocol... Addgene Protocols On this page you'll find practical lab protocols that you can use for ...A wide variety of protocols from Addgene that can be used for basic molecular biology, plasmid cloning...range of applications, with videos for select protocols in the right-hand column. You can find even more...Addgene Video Page . We hope that you'll find these protocols useful in your own work. Please feel free to email...materials on a balance Basic Molecular Biology These protocols are the building blocks for many more complicated...Quantification Measure DNA concentration with a spectrophotometer Restriction Digests Cut and analyze plasmid...Video! Polymerase Chain Reaction (PCR) Basic PCR protocol with tips and FAQ Watch the Video! Purifying DNA... -
Molecular Biology Protocol - Restriction Digest of Plasmid DNA
TypeProtocol...Protocol, tips, and FAQ for how to perform a restriction digest of plasmid DNA... Protocols Restriction ...diagnostic digest . Last Upload: Oct. 11th, 2016 Protocol Video Watch the video below to learn how to analyze... -
Protocol - How to Purify DNA from an Agarose Gel
TypeProtocol...based cloning. Protocol: Gel Purification Follow the Agarose Gel Electrophoresis Protocol with the following...Protocol for how to purify DNA from an agarose gel, including tips to help improve the resolution and... Protocols Purifying DNA from an Agarose Gel Purifying DNA from an Agarose Gel ... -
Video Library
TypeProtocol...video protocols, tips on using Addgene's resources, and career advice. For written lab protocol material...questions. Protocols Video protocol guides for standard laboratory procedures. (Link opens in a new window...including how-to screen captures, lab procedure protocols, and career advice videos. Educational...Resources Addgene Videos Addgene Videos Protocols How-To Videos Career Videos Browse Addgene's collection of...material, visit our Addgene Protocols page . Please feel free to email us at [email protected] with any questions...culture bacteria in the lab Pouring LB Agar Plates Protocol Streaking Bacteria on Plates Isolate single bacterial... on an agar plate Streaking Bacteria on Plates Protocol Diagnostic Restriction Digest Verify a plasmid... -
Kit Free RNA Extraction
TypeProtocol...Procedure Option #1 - Solution D Protocol Before starting this protocol, make sure to prepare solution ...and storing those in -80°C. Solution D Protocol TRIzol® Protocol Top...Follow this protocol to perform RNA extraction without a kit. Protocols... Protocols RNA Extraction Without a Kit ... Molecular Biology Reference Introduction This protocol describes how to extract RNA from tissues or cells...cells without a kit. The general steps of this protocol include homogenization/lysis of cells or tissues... of RNA, precipitation, and resuspension. This protocol provides two options: using Solution D (see reagent... -
Affinity Purification of Recombinant Antibodies with Protein A or Protein G
TypeProtocol...Antibodies Immunocytochemistry Protocol Western Blot Protocol Introduction This protocol describes how to affinity...outcomes when using these protocols. However, please be aware that the protocol may need to be adjusted ...This protocol describes how to affinity purify recombinant antibodies from cell culture supernatant using... Protocols Recombinant Antibody Affinity Purification with Protein... believe that sharing the full details of our protocols supports reproducibility and accelerates science...waste procedures. Last Update: February 7, 2022 Protocol Video Watch this instructional video to learn ...compatible with 50 mL conical tubes NanoDrop spectrophotometer 37 °C, 5% CO 2 incubator with shaking platform... -
CRISPR Library Amplification
TypeProtocol...specific protocol if one is available. If a pooled library does not yet have an amplification protocol, the...the following protocol can be used as a starting point for CRISPR libraries. This protocol allows the amplification...these protocols can be found on the Pooled Library page for the specific library. This protocol assumes... our protocol on Bacterial Transformation . The generalized pooled library amplification protocol is expected...Follow this protocol to perform amplification of CRISPR pooled plasmid libraries in Escherichia coli ... Protocols CRISPR Library Amplification CRISPR Library Amplification You may also like... Pooled...pooled library material pages for amplification protocols that have been developed by the depositor for ... -
Antibody Validation Using the Indirect ELISA Method
TypeProtocol...Immunocytochemistry Protocol Recombinant Antibody Purification Protocol Antibody Guide Introduction This protocol describes...outcomes when using these protocols. However, please be aware that the protocol may need to be adjusted ...of each well at 450 nm on the spectrophotometer. Use the spectrophotometer to generate a standard curve...This protocol describes how to run an indirect ELISA against a purified antigen to demonstrate the expected... Protocols Antibody Validation Using the Indirect ELISA Method Antibody...purified antigen to validate an antibody. This protocol will describe how to create a standard curve using... believe that sharing the full details of our protocols supports reproducibility and accelerates science... -
Isolating a Monoclonal Cell Population by Limiting Dilution
TypeProtocol... Virus Blog Posts Addgene Protocols Viral Service Introduction This protocol describes how to generate...Workflow Timeline This protocol begins with a stable cell pool. For details, see our protocol for generating ...This protocol describes how to generate a monoclonal cell line from a polyclonal pool of stable cells... Protocols Cloning by Limiting Dilution Isolating a Monoclonal Cell Population by Limiting Dilution... at 4 °C. Polyclonal stable cell pool: see our protocol for generating stable cell lines with lentivirus...to grow under the conditions described in this protocol. Conditioned medium (details below) can be used...monitored regularly for transgene expression. This protocol has been optimized for adherent cells but can ... -
Centrifugation
TypeProtocol... Protocols Centrifugation Centrifugation Protocol Intro to the Lab Bench Check...Check out more protocols and videos to help you get started in the lab! Introduction A centrifuge is a ...or keep samples at specific temperatures. This protocol will cover the general procedure and features ... will have essentially the same components. The photo to the left shows a front view of an open tabletop...tabletop microcentrifuge with the lid open, the rotor with microfuge tubes in place, and the control panel...centrifuge, with tubes evenly distributed around the rotor. How to Centrifuge Now that you know the different...working smoothly. Place your sample tubes into the rotor so that they are distributed evenly around the center... -
AAV Purification by Iodixanol Gradient Ultracentrifugation
TypeProtocol... Guides Virus Blog Posts Mol Bio Protocols Introduction This protocol can be used to purify AAV of any...a new window) from Sigma-Aldrich. Protocol Video Watch the protocol video below to learn setup and use...Follow this protocol using Iodixanol gradient ultracentrifugation to get pure adeno-associated virus ... Protocols AAV Purification by Iodixanol Gradient Ultracentrifugation...purified AAV in vivo . The iodixanol gradient in this protocol is composed of steps that separate out contaminants... one (long) day. Equipment Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep (60% Iodixanol)... Centrifuge at 350,000 x g for 90 min in a T70i rotor at 10 °C. Pro-Tip If you need more time, you can... -
Handling Plasmids from Addgene - Purifying Plasmid DNA
TypeProtocol...from a bacterial culture, including protocols, tips, and FAQ... Protocols Recovering Plasmid DNA from Bacterial...concentrations ranging from 150 ng/μL to several μg/μL. The protocol below is meant to describe the general procedure...purification without using a kit, you can find a protocol for kit-free plasmid mini-prep at the bottom of... 70% ethanol TE buffer 3 M Na-acetate (pH 4.8) Protocol: Generalized DNA Purification Grow an overnight...bacteria . Pro-Tip Refer to appropriate DNA prep protocol for volume of bacteria to grow (low copy plasmids... DNA concentration and purity click here . Top Protocol: Kit-free Alkaline Lysis Plasmid Miniprep Prepare...