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We narrowed to 401 results for: PREP

Showing: 181 - 200 of 401 results

  1. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...Resources on Addgene.org Find ready-to-use AAV Preps Find AAV plasmids Browse all viral vectors ...intracellular transport machinery. Addgene offers viral preparations of AAV2-retro vectors with a variety of transgene...

  3. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...cloning, and titering and testing your viral preparations. ... Video! Inoculating a Liquid Bacterial Culture Prepare and grow bacteria in liquid medium Watch the Video...long term use Watch the Video! DNA Purification Miniprep, phenol-chloroform extract, and precipitate DNA...Protocols for titering and testing your virus preparations. Name Description Link to Video General Transfection...Ultracentrifugation Purify adeno-associated virus from a preparation produced in mammalian cells Watch the Video! ...physical titer of your adeno-associated virus preparation Watch the Video! AAV Titration by ddPCR Use ddPCR...physical titer of your adeno-associated virus preparation Lentivirus Titration by ddPCR Use ddPCR to measure...

  4. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...able to start an overnight culture for plasmid DNA prep the following day. Tips and FAQ The optimal concentration...glycerol stock before you begin your plasmid mini-prep. Try not to freeze/thaw your glycerol stock too ...15-25% glycerol. You can prepare the glycerol stock the same time you prepare your plasmid DNA. In the...

  5. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ... do a mini-prep you will usually want to start 2 mL in a falcon tube, but for larger preps you might want...inoculate bacteria in liquid culture. Protocol Prepare liquid LB. For example, to make 400 mL of LB, weigh...

  6. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...the prep with full (genome containing) particles. On average, iodixanol-purified AAV preparations contain...ultracentrifugation to get pure adeno-associated virus (AAV) preps for use in your research. Protocols...Ultracentrifuge T70i rotor or equivalent Reagents Opti-Prep (60% Iodixanol) QuickSeal tubes QuickSeal tube spacers...separate out contaminants from an impure AAV preparation. The 15% iodixanol step has 1M NaCl to destabilize...iodixanol solution is available under the name OptiPrep (Link opens in a new window) from Sigma-Aldrich...Centrifugal filter units (MWCO 100 kDa) Reagent Preparation 1 M NaCl/PBS-MK buffer Dissolve 5.84 g of NaCl... and 2 mL of 5 M NaCl to 43 mL PBS Procedure Preparation and loading of the iodixanol gradient: 15% iodixanol...

  7. Fluorescence Titering Assay

    Type
    Protocol
    ...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...conical tubes, VWR 21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM...Protocol that can be used to titer lentiviral preparations carrying a fluorescence marker. Protocols...This protocol can be used to titer lentiviral preparations that carry a fluorescence marker. Determining... 75,000 cells into each well of a 6-well dish. Prepare a batch of cells as follows: Dilute 525,000 cells...thaw the lentiviral aliquot on ice prior to use. Prepare dilutions of the lentivirus into DMEM complete ...

  9. Colony Formation Titering Assay

    Type
    Protocol
    ...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...conical tubes, VWR 21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM...This protocol can be used to titer lentiviral preparations that confer antibiotic resistance. Protocols...This protocol can be used to titer lentivirus preparations that confer antibiotic resistance. Determining...Determining the titer of your lentiviral preparation allows you to control the multiplicity of infection (MOI... Use this dose for the colony formation assay. Prepare a batch of DMEM complete containing 10 μg/mL polybrene...thaw the lentiviral aliquot on ice prior to use. Prepare 10-fold serial dilutions of the lentivirus into...

  10. DNA Quantification

    Type
    Protocol
    ...NanoDrop to measure your samples, place 1-2µL of mini-prepped DNA onto the pedestal. Close the lid and click ...

  11. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously filtered) Microcentrifuge...15 mL conical tubes, VWR 21008-216 Lentivirus preparation Appropriate antibiotic for selection (e.g. puromycin...puromycin, blasticidin) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine (...determine the lowest dose that kills all of the cells. Prepare a batch of DMEM complete + 10 µg/mL polybrene by...Thaw the lentiviral aliquot on ice prior to use. Prepare a range of dilutions of the lentivirus in DMEM ...cell solution in this step. To seed the cells: Prepare a batch of cells as follows: Dilute 350,000 cells...growing well and have been sufficiently expanded, prepare cell stocks and/or harvest to test for protein ...

  12. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...you can find a protocol for kit-free plasmid mini-prep at the bottom of this page. Last Update: Feb. 8,...culture of bacteria . Pro-Tip Refer to appropriate DNA prep protocol for volume of bacteria to grow (low copy... Protocol: Kit-free Alkaline Lysis Plasmid Miniprep Prepare the following solutions: Solution I - Resuspension...pellet the bacteria before proceeding with DNA preparation. Pro-Tip If your entire overnight culture cannot...

  13. Plan Your Experiment

    Type
    Guide
    ...most common. Using viral vectors takes a bit more prep work, as you first have to produce the viral particles...Browse Addgene's collection of in-stock lentiviral preps . AAV vectors AAV vectors offer some advantages ...size limitation . Browse Addgene's in-stock AAV preps or try Addgene's Packaged on Request service. Delivery...

  14. Antibody Guide

    Type
    Collection
    ...conjugating them in the lab? How will the sample be prepped, and will this or other factors affect epitope ...isotype. Overview of application Fix your samples. Prepare your samples using permeabilization, antigen retrieval...and controls follow that of IF (above). Sample preparation, signaling excitation, and assay readout vary...

  15. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...appropriate imaging system. Figure 1 Recombinant antibody preps should have 2 clear bands at ~50 kDa and ~25 kDa... Pro-Tip Addgene only uses recombinant antibody preps that have a purity of ≥90% but some labs use unpurified...prestained protein ladder on ice. Reagent Preparation Prepare IgG Standard Dilute the 2.5 mg/mL IgG isotype.... Add 8 µL of PBS to a total volume of 13 µL . Prepare the recombinant antibody Dilute or concentrate ...microfuge tube. Add 8 µL of PBS to total 13 µL . Prepare 1X MOPS buffer Dilute 25 mL of 20X MOPS buffer ...

  16. Lentiviral Vector Guide

    Type
    Guide
    ...Plasmids Collection In-Stock Lentiviral Preps Addgene's Viral Prep Service Viral Vectors 101 eBook Addgene's...Browse Addgene's collection of in-stock lentiviral preps . CRISPR Genomic Screens Genome-wide screens are...

  17. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Packaging Services Packaged-on-Request AAV Preps In-Stock AAV Preps Resources on addgene.org AAV Plasmids ...more information. Browse Addgene’s in-stock viral preps . AAV retrograde (AAVrg) Directed evolution has ...were discovered as a contaminant of adenovirus preparations, which is how they got their name, but they ...

  18. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... number of genome-containing particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s... number of genome-containing particles of an AAV prep using SYBR green technology. This protocol was validated...each step Reagent Preparation Master Mix: Count the number of samples (n) and prepare master mix for an...μL Nuclease Free Water 4.7 μL 470 μL Procedure Prepare a plasmid stock of 2 x 10 9 molecules/μL to generate...

  20. We're Updating Our Hold for Publication Policy

    Type
    Blog Post
    ...unpublished materials. What about preprints? Addgene supports preprints and encourages authors to share ...alongside a journal article, a preprint, or without any paper at all. Posting preprints and openly sharing materials...reproducibility, and accelerate science. We encourage preprint authors to deposit materials without a hold or...or to release the hold as soon as the preprint is posted in order to support these goals. And we can update...article version any time later. We recognize that a preprint isn’t necessarily a clear sign that you are ready...your plasmids, so we won’t immediately release preprint-associated materials the way we will with journal...to consider sharing them openly as soon as that preprint is online!   Thanks for reading, and for depositing...
Showing: 181 - 200 of 401 results