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

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We narrowed to 994 results for: Ran

Showing: 941 - 960 of 994 results

  4. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...recipient plasmid backbone. Transformation Proceed with the transformation according to the manufacturer...from your transformation will give you the first indication as to whether your transformation worked. Your...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary PCR based cloning is incredibly versatile...chance for mis-priming, so you can use a pretty wide range of annealing temperatures, but you may need to increase...competent cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent...conduct a positive control to ensure that your transformation worked. You could also try varying the amount...cloning. DNA replication by PCR has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million...

  6. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...self-ligating recipient plasmid backbone. Transformation Transform your ligation reaction into your bacterial...from your transformation will give you the first indication as to whether your transformation worked. our...Digest of Plasmid DNA DNA Ligation Bacterial Transformation Summary The following technique can be used...competent cells. For most standard cloning, you can transform 1-2μl of your ligation reaction into competent...conduct a positive control to ensure that your transformation worked. You should also verify that you are...

  7. Ligation Independent Cloning

    Type
    Protocol
    ...now ready for transformation. Use 1-2 μl of annealing reaction for each transformation, following a standard...stability to hold the plasmid together through the transformation/replication process. LIC employs long overhangs...association between fragments, allowing for transformation without ligation. Because of its dual polymerase...same principle (homologous to the first G on 3'-5' strand from cut site). For simplicity, only the 5' primer...heating to 75° for 20 minutes. Step 6: Anneal and Transform Mix your treated vector and insert at a molar ...

  8. Video Library

    Type
    Protocol
    ... tips Bacterial Transformations We walk you through a standard plasmid transformation protocol and offer...tricks to reduce transformation time and increase efficiency Bacterial Transformation Protocol Inoculating...when requesting samples from Addgene Technology Transfer How to Order A step-by-step walkthrough of Addgene's...Addgene's Business Development Manager, discusses his transition from the Scientist Team to Business Team, his...

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

    Type
    Protocol
    ...practical lab protocols that you can use for a wide range of applications, with videos for select protocols...Ligation Assemble DNA using DNA ligase Bacterial Transformation Introduce DNA into a bacterial strain Watch...preparations. Name Description Link to Video General Transfection Introduce plasmid DNA to mammalian cells Lentivirus...Produce lentivirus with a polyethyenimine (PEI) transfection protocol Fluorescence Titering Assay for Lentivirus...applications. Name Description Link to Video Transfection Introduce plasmid DNA to mammalian cells to ...

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...
Showing: 941 - 960 of 994 results