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Showing: 601 - 620 of 749 results
  1. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...how to design primers for cloning purposes. What do I do if my PCR isn't working? Try adding 1µl of 25mM...36.8 μL Sterile dH 2 O (variable) Note: If you are doing multiple PCRs, save time by creating a “master mix...added to the master mix. Forward and Reverse Primers DO NOT get added to a master mix. Place reaction tubes...reverse primers to the thinned walled PCR tubes. Note: Do this before adding the master mix so you know that...according to the length of the target sequence. FAQ How do I design primers? See our reference page on how to...The ideal setup for this troubleshooting step is to do one reaction with each, and one reaction with both...templates. What does each ingredient specifically do? Template DNA: Contains the portion of DNA that we...
  2. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ... provided you with a few ideas to consider if you do run into a roadblock and encourage you to check out...overcome this issue, we recommend using a range of doses of lentivirus when transducing a target cell line...transduction experiment. Media choice, reaction vessel, dose of virus, volume and timing of transduction will...
  3. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...Despite their advantages, mammalian expressions systems do require demanding cell culture conditions compared...
  4. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...been used to treat 10 infants with this disease. To do so, their T-cells were grown in vitro, their mutations...to be untreatable until 2008. Following a single dose of subretinal administration of rAAV2 carrying the...
  5. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...transformation . Tips and FAQ Do controls When doing ligations you should ALWAYS do a vector alone + ligase ...you know has only been thawed once before. Always do controls. See Tips and FAQ below for details. Try...situations where the 3:1 ratio is not working or when doing more complicated cloning. While 3:1 will get you...
  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...plasmid tools that have come through our doors in 2015. Do you have a favorite new plasmid tool from the past... are turned on or off in a given area and the can do so in a very specific, precise manner. Previously... are bound to the coding region are displaced and do not rebind because they are transported back to the...their quest to understand lncRNAs: What can lncRNAs do by themselves? The Rinn lab has developed a novel...regulation has roles in many cellular functions, from endosomal trafficking to signaling pathways to the cell ...reticulum (ER), mitochondria, cytoskeleton, and endosomes. These fusion proteins can act as markers for ...Microtubules mCherry-alpha-tubulin Early Endosome Vacuolar Compartment mCherry-Rab5 BFP-Rab5 ...
  7. CRISPR Library Amplification

    Type
    Protocol
    ...total plasmid DNA. Tips and Troubleshooting What do I do if my transformation efficiency is not high enough... Pooled Libraries Molecular Biology Reference How do I process my Addgene pooled library? Introduction...Endura from two separate transformations). Pro-Tip Do not pipette repeatedly or mix when removing SOC containing...Critical Be careful not to rip or shred the agar. Do so by gentle spreading. Some spreaders have a sharp...Maxi Kit (one conical is its own Maxiprep). Critical Do not freeze pellets for later purification. Immediately...colder reagents (not including the recovery media!). Do not proceed with Maxipreps or NGS until adequate ...representation is maintained. Maxipreps - Less is more: Do not overload the Maxipreps as yield can dramatically...
  8. Plasmids 101: Degron Tags

    Type
    Blog Post
    ... protein abundance. Finally, some degron tags can do more than just degrade! Read on to find out more....
  9. Hot Plasmids February 2024

    Type
    Blog Post
    ...01756-1. PMID: 37095348. Breaking up is hard to do: Monomeric StayGold variants By Mike Lacy In 2022... it with an “assist peptide” that facilitates endosomal escape. Combining Cas9-PAGE with lentiviral sgRNA...based vectors (and vice versa), enabling repeat dosing. In a proof-of-concept study, mouse LY6A was delivered...
Showing: 601 - 620 of 749 results