Skip to main content
Addgene
Open Menu

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 1,010 results for: RING;

Showing: 971 - 980 of 1010 results

  1. AAV Production in HEK293 Cells

    Type
    Protocol
    ...~7.4. Autoclave or sterile filter. Pro-Tip Stirring during the cooling period is recommended or the solution...glucose, lower FBS media causes less filter clogging during harvesting. The addition of sorbitol has been shown...PEI powder into 100 mL of deionized water. While stirring, slowly add hydrochloric acid until the solution... challenging due to the high viscosity of PEG. Stirring under medium heat will promote faster dissolution...overgrow your cells. Pass the cells twice a week during the maintenance phase and do not allow cells to... °C for 1 h, then keep at 4 °C for 3 h without stirring to allow full precipitation. Precipitation of ...

  2. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ... of reagents and a basic heating and cooling (denaturing and annealing) cycle. The process became automated...in an appropriate buffer, a series of heating (denaturing) and cooling (annealing) steps allow the Taq ...template strand to the point where the strands start denaturing and the hydrogen bonds are broken between the... tops to the PCR tubes, gently tap each tube to bring all the liquid to the bottom before placing it in...sequence is GC-rich, increase the time of the denaturing. Your 5’ and 3’ primers should be designed to... that can withstand radical temperature changes during a typical PCR. The DNA polymerase has an optimum...

  3. Pipetting Protocol

    Type
    Protocol
    ...pipette shows the volume adjustment ring right below the plunger. This ring changes the pipette volume. Below... volume for the the pipette, and the tip ejector ring at the bottom which pushes the pipette tip off of...tip is so that the same pipette can be used for measuring different samples without cross contamination ...your fingers to avoid contaminating the tip or puncturing your glove. Ensure that the tip is properly set...

  4. AAV ddPCR Titration

    Type
    Protocol
    ... particular sample. For additional tips on AAV titering using ddPCR, read this blog post on ddPCR for ...all the way, just ensure the samples are covered. Bring the PCR tubes to the BSC used for dilutions. Without...Lightly cap the PCR tubes. Generate the Droplets Bring the PCR tubes to the droplet generation BSC. Without...samples on ice or pre-chilled 96-well freezer blocks during use. In this protocol, a dilution series is prepared...dilutions and titrations For additional tips on AAV titering using ddPCR, read our blog post. Last reviewed...

  5. DNA Quantification

    Type
    Protocol
    ...like... Inoculating a Liquid Bacterial Culture Recovering Plasmid DNA from Bacterial Culture Restriction...Restriction Digest of Plasmid DNA Background Information During several different stages of molecular cloning, ...specific wavelength that they maximally absorb. By measuring the absorbance of a liquid you can accurately ...specific to your lab. Spectrophotometer Tips Before measuring any samples, be sure to ‘blank’ the spectrophotometer...

  6. CRISPR Library Amplification

    Type
    Protocol
    ...as possible due to the inherent possibility of altering the composition of the library. Bottlenecks, fitness...representation of individual plasmids in the pooled library during amplification. This protocol is designed to be ...pulling motion Take care not to split or gouge agar during the scraping process. Add each scrape into a 50...efficiency! Ensure that no arcing is taking place during electroporation. Arcing would manifest as a loud... often accompanied by some light in the cuvette during the electroporation step. I got a heavy recombined...

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

    Type
    Protocol
    ... basic molecular biology, plasmid cloning, and titering and testing your viral preparations. ...targeting human and mouse genes. Virus Protocols for titering and testing your virus preparations. Name Description...polyethyenimine (PEI) transfection protocol Fluorescence Titering Assay for Lentivirus Quantify virus using fluorescence...fluorescence measurements Colony Formation Titering Assay for Lentivirus Quantify virus by counting antibiotic...

  8. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...QuickSeal tube spacers 16 ga needle 18 ga needle 10 mL syringe 18 ga blunt edge needles, Hamilton 1X PBS pH 7.4...QuickSeal tube in the order below using a 10 mL syringe and a blunt edge 18 ga Hamilton needle, taking ...interface with an 18 ga needle attached to a 10 mL syringe. The bevel of the needle should be up, facing the... If the concentrate volume is less than 500 µL, bring up the volume with formulation buffer. Use a P1000...

  9. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...loading buffer, New England Biolabs B7022S Procedure Pouring a Standard 1% Agarose Gel: Measure 1 g of agarose...the solution heats up.). Caution HOT! Be careful stirring, eruptive boiling can occur. Pro-Tip It is a good...on the sample to prevent bubbles or buffer from entering the tip. Place the very top of the tip of the ...Note: When using UV light, protect your skin by wearing safety goggles or a face shield, gloves and a lab...

  10. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...Protein A or Protein G columns. Sharing speeds science. We believe that sharing the full details of our protocols...89039-656 Microcentrifuge tubes, VWR 89126-724 Filtering system 0.2 µm PES, VWR 431098 Sodium phosphate...filtration unit, 500 mL, rapid-flow, VWR 73520-984 Syringes, 30 mL, VWR BD302832 Filter, 0.2 µm (luer-lock...
Showing: 971 - 980 of 1010 results