Skip to main content
Addgene
Showing: 61 - 80 of 290 results
  1. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...join any fragments end-to-end without the exo/polymerase generation of annealed complementary regions....and even include profiling single nucleotide polymorphisms (SNPs) (5,12,13). For example, in the Ligase...Shuman, Stewart, and Christopher D. Lima. "The polynucleotide ligase and RNA capping enzyme superfamily of...for characterization of resistance-mediating polymorphisms in African samples of Plasmodium falciparum....
  2. SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture

    Type
    Blog Post
    ... uses do you foresee for this tool? Howarth: Polyproteins are useful for mechanical study of proteins ...proteins by atomic force microscopy (AFM). Maybe polyantibodies could help in the capture of other rare cell ...References: SpyLigase peptide-peptide ligation polymerizes affibodies to enhance magnetic cancer cell capture...
  3. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... a duplicated SV40 polyadenylation region instead of the synthetic polyadenylation signal present in pGL3...recruits up to 24 copies of a protein to a target polypeptide. Multimerization in the SunTag system occurs ... simplifies the process of creating complex polycistronic lentiviruses. The MuLE toolbox contains a series...constitutive promoters, 2 includible promoters, 3 polyA terminators and various pieces for inducible expression...UAAs via a re-assigned stop codon (TAG). This polyspecificity enables the site-specific introduction of many...
  4. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    ...microelectronic fibers made from thermoplastic polymers that can work with all the bells and whistles....called thermal drawing, essentially where blocks of polymers are heated up and pulled to a desired thickness... small molecules.   Fun fact: A 1 cm block of polymer can generate many meters of fibers!     ...
  5. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ... bodies are a polymer of many copies of monomeric subunits. Unlike cytoskeletal polymers, however, R bodies...force by dynamically rearranging into different polymerization states. Instead, R body monomers respond to...
  6. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ... PCR and surveyor. I used DNA polymerase Takara Ex Taq ™ DNA Polymerase for my genomic PCR, but couldn't...In our hands, Herculase II Fusion polymerase or Kapa Hifi Polymerase work very well. Some groups have ...your HR by doing Restriction Fragment Length Polymorphism (RFLP) ( see Figure 4 of the Cong, et. al, 2013...
  7. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ...Marchantia polymorpha MpU6 none S. pyogenes Hara-Nishimura pMpGE_En03 71535 Other/Marchantia polymorpha MpU6...pyogenes Ventura pCRISPRyl 70007 Other/Yarrowia lipolytica SCR1'-tRNA yes, cut S. pyogenes LEU2 Wheeldon...pyogenes Draetta pCAS1yl 73226 Other/Yarrowia lipolytica yes, cut S. pyogenes LEU2 Yang pLCKO 73311 Mammalian...Gateway recombination-based cloning of complex polycistronic lentiviruses. Can express Cas9 with up to 3 ...in a single vector. sgRNAs are expressed as polycistronic glycine tRNA-gRNA genes (PTGs) and then PTG ...
  8. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...Ligation Buffer, 6.5 μl ddH 2 O, and 0.5 μl T4 Polynucleotide Kinase (PNK) (10,000 U/ml). NOTE: Phosphorylated...acceptable. The bulk sorted cells are composed of a polyclonal population exposed to sgRNA-A and sgRNA-B (see... band” in separate reactions. NOTE: Numerous polymerases may be used for step 6.3 . Run samples in a thermocycler...a single reaction. Optimize multiplexing in a polyclonal population ( i.e., bulk sorted cells) before ...biallelic deletion clones with a proofreading polymerase and clone the amplicons ( e.g., with a PCR cloning...
  9. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...Addgene blog   Droplet Digital PCR for AAV QuantificationPolymerase Chain Reaction Overview and Applications...Applications Resources on Addgene.org Protocol: Polymerase Chain Reaction (PCR) Protocol: How to Design a primer...SYBR Green I and TaqMan quantitative real-time polymerase chain reaction methods for the determination ...
  10. Plasmids 101: Protein Expression

    Type
    Blog Post
    ... can then be translated at the ribosome into polypeptide chains that make up the primary structure of ...purified for use in molecular biology research (eg. polymerases and other enzymes might be purified and used ...translation machinery. These include ribosomes, RNA polymerase, tRNAs, ribonucleotides and amino acids. Cell-free...
  11. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...fragments are then combined with an exonuclease, a polymerase, and a DNA ligase to carry out a pooled set of...topoisomerase I cut site, toposisomerase I, and a taq polymerase-generated insert. When mixed, topoisomerase I... could also be a great choice, because the T4 polymerase needed for this method is relatively inexpensive...
  12. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...set up a standard PCR reaction (primers, dNTPs, polymerase) using the supernatant of lysed bacteria as template...standard PCR reaction: combine template, primers, polymerase, and dNTPs and then incubate with a standard ...heating step of the PCR reaction. A standard Taq polymerase is sufficient. Controls: Controls can make ...
  13. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ... separate plasmid (envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein... be disposed of following your lab’s biohazard policies. Administering Virus Many virus experiments involve... accident-prone researchers. Note that safety policies will vary slightly from organization to organization...with your specific organization/department/lab’s policies – basically, you’ll need to get trained in each...
  14. Viral Production at Addgene

    Type
    Blog Post
    ...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...centrifugation and then by filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then ...lentiviral preparation by precipitation in 10% polyethylene glycol (PEG) followed by centrifugation. Precipitated...
  15. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...with a “sloppy” version of PCR that results in a polymerase error rate of up to 2% per nucleotide position...modifications: 1) increased concentration of Taq polymerase, 2) increased PCR extension time, 3) increased...degenerate primers and the high-fidelity Phusion polymerase are used to introduce point mutations into the...
  16. Adeno-associated virus (AAV) Plasmids

    Type
    Collection
    ...production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-1 cap gene in Sf9 insect cells Kotin 65214...production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-2 cap gene in Sf9 insect cells Kotin 65215...production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-6 cap gene in Sf9 insect cells Kotin 65216...production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-8 cap gene in Sf9 insect cells Kotin 92307...
  17. Viral Production

    Type
    Collection
    ...Samples of viral preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver...centrifugation and then by filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then ... 293T cell line in the presence of 10.0 μg/mL polybrene. This ensures that any loss of titer associated...
  18. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...biorender.com.    Since antibodies, especially polyclonal antibodies, can vary lot-to-lot, this range needs...this starting point. You can also consider using polyclonal or recombinant multiclonal antibodies to increase... time Not enough antibody binding sites Polyclonal or recombinant multiclonal antibodies Protein...
Showing: 61 - 80 of 290 results