Skip to main content
Addgene

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

Information

In Category

We narrowed to 45 results for: Ran

Showing: 21 - 30 of 45 results

  1. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...Restriction Digest of Plasmid DNA Bacterial Transformation Background Information The final step in the...the backbone and the complete plasmid can be transformed into bacterial cells for propagation. The majority...recognition sequence, which results in a single stranded overhang on the digested end of the DNA fragment... incubation at 37°C. Proceed with bacterial transformation . Tips and FAQ Do controls When doing ligations...contamination of intact plasmid in ligation or transformation reagents Optimizing the Vector:Insert Ratio...

  2. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies Recombinant Antibody... Inclusion of this information is solely for transparency intended to support reproducibility in science....125 mg/mL: 10 µL of 0.25 mg/mL with 10 µL PBS Transfer 5 µL of each standard to a microfuge tube. Add... antibody to a concentration of 0.9–1.1 mg/mL. Transfer 5 µL of recombinant antibody to a microfuge tube... menu and select Scatter chart. Change the data range and select the cells where the x-axis is represented...

  3. Weighing Reagents Protocol

    Type
    Protocol
    ...material that you are weighing by looking for a weight range on the scale. Make sure that the weight of the material... material you’re weighing out is within this range. If you need less than a gram of material, use an analytical...cross-contamination with other substances. They also help you transfer the material to your tube or container. Pro-Tip...weighed out the correct amount of your reagent, transfer it into your container. Make sure that any spills...

  4. Gibson Assembly Protocol

    Type
    Protocol
    ...of a Gibson Assembly is a fully ligated double-stranded DNA molecule. This has proven to be an efficient...for the reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing back from the DNA 5...fragment. Pro-Tip Add Extreme Thermostable Single-Stranded DNA-Binding protein (ET SSB) to the isothermal...You can purchase master mix or make your own. Transform bacteria with the DNA and screen for the correct...

  5. Immunocytochemistry

    Type
    Protocol
    ... Inclusion of this information is solely for transparency intended to support reproducibility in science... antibody concentration will vary but generally ranges from 1-10 µg/mL. Add 500 µL of the diluted antibody... antibody concentration will vary but generally ranges from 1-10 µg/mL. Add 500 µL fluorescently-labeled...you know expresses the protein, such as cells transfected with a plasmid to express the protein of interest...

  6. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...Bacterial Culture You may also like... Bacterial Transformation Agarose Gel Electrophoresis Agarose Gel DNA...much as several miligrams and at concentrations ranging from 150 ng/μL to several μg/μL. The protocol below...optional) Reagents Overnight culture of bacteria transformed with your plasmid Resuspension buffer Denaturing... a pancreatic ribonuclease that digests single-stranded RNA. Optional: Perform phenol-chloroform extraction...

  7. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...sample (do not vortex) Incubate 30 min at 37 °C Transfer to ice ** Critical: do NOT treat your plasmid ...dilutions 5–8 Note: at Addgene, samples typically range from 1 x 10 12 GC/mL to >2 x 10 13 GC/mL and we ... and forth at least 5 times. Seal plate with transparent film. Centrifuge at 3,000 rpm for 2 min to bring...) ~ 1.0, E (efficiency of PCR) ~100% (90%–110% range is acceptable) Baseline removal: all samples will...

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

    Type
    Protocol
    ...Collection Antibody Blog Posts Antibody Guide Transfection for Recombinant Antibodies Immunocytochemistry... Inclusion of this information is solely for transparency intended to support reproducibility in science...recombinant antibody to room temperature (see Transfection for Recombinant Antibody protocol ). Pro-Tip... has reached the desired concentration. Gently transfer the sample from the reservoir to a LoBind tube...

  9. AAV ddPCR Titration

    Type
    Protocol
    ...outlined in this protocol are based on an AAV titer range of 5E+12–5E+13 GC/mL, where GC is genome copies ...the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet generator....while slowly and gently aspirating the droplets. Transfer the droplets to a prechilled PCR plate. Pro-Tip... 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open the QuantaSoft...

  10. Protocol - How to Design Primers

    Type
    Protocol
    ...complementary to template strand). However, primers do not need to correspond to the template strand completely; it...primer corresponds completely to the template DNA strand so elongation can proceed. Usually a guanine or...
Showing: 21 - 30 of 45 results