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Showing: 1 - 16 of 16 results
  1. Antibody Guide

    Type
    Guide
    ...reaction. Many antibodies can be purchased already conjugated, which can save significant time in the lab. However...choice, you can conjugate antibodies in the lab. The complexity of this can vary significantly based on ... B cell can clone itself to enable higher production of a single antibody, but each B cell can only produce... and can be used to increase flexibility or specificity in antibody-based methods. Antibodies can recognize...antibodies can be isotype-specific or even isotype subclass-specific. This specificity can be used to ...that you can detect in a single assay is limited by the number of signaling molecules that can be distinguished...may be present in. However, polyclonals can vary significantly from lot to lot, as immune responses have...
  2. Molecular Cloning Techniques

    Type
    Guide
    ... enzyme target sites can be repeated throughout a specific DNA sequence. This can make it difficult at...no homology ends can still be fused in a seamless manner. To accomplish this, you can transform into yeast...and discovery. With molecular cloning, scientists can amplify and manipulate genes of interest and then...complementary overhangs, or which are both blunt-ended, can then be fused during a ligation reaction with T4 ...usually within the multiple cloning site (MCS). You can easily move (subclone) any piece of DNA that already...leaves behind a short scar in the DNA sequence and can be time-consuming compared to other cloning methods...’ and 3’ ends of the DNA sequence. This fragment can then be cloned into a Gateway donor plasmid, which...
  3. Molecular Biology Reference

    Type
    Guide
    ... information and can replicate independently from chromosomal DNA. Although they can be found in archaea... and can be specific to a certain species, tissue, or cell type. The strength of the promoter can control...every protein within an organism. Before proteins can be translated, the DNA is converted to RNA in a process... only 20 natural amino acids, so multiple codons can encode the same amino acid — a phenomenon called ... to the host, such as antibiotic resistance, and can be passed from one bacterium to another by a type...recombinant DNA, in which many identical DNA fragments can be generated. The combination of a DNA fragment with...backbone generates a recombinant DNA molecule, which can be used to study DNA fragments of interest, such ...
  4. Plan Your Experiment

    Type
    Guide
    ...transcriptase (RT)) Can edit within a wider window than most base editors Editing efficiency can be low, but ...within the gene itself can also be effective, as the Cas9 bound to the DNA can interfere with transcription...transfection or electroporation. Expression can be transient, or you can generate stable cell lines if your plasmid...targeted edits can lead to changes in more than just the DNA sequence. If this is the case, you can confirm ...different Cas proteins available, and researchers can adapt these experiments for different organisms. ... to have a biological question in mind. Then you can decide how to answer that question using genetic ...but generally higher than HDR pegRNA optimization can be challenging Prime Edit Repress or Interfere (CRISPRi...
  5. Lentiviral Vector Guide

    Type
    Guide
    ...gamma-retroviruses can only infect mitotically active cell types. This means that lentiviruses can infect a greater...Plasmid Can be packaged ONLY by a second-generation packaging system that includes the tat gene Can be packaged...continue their lifecycle and replicate. This feature can be harnessed for stable delivery of various genes...have been engineered into lentiviral vectors that can be safely used in laboratory settings. These engineered...variety of cell types and models. Lentiviral vectors can target non-dividing cells, enable stable long-term...split into two plasmids. Although safer, this system can be more cumbersome to use and result in lower viral... contains env Third-generation transfer plasmids can be packaged by either second- or third-generation...
  6. CRISPR Guide

    Type
    Guide
    ...monocytogenes , which can inhibit the CRISPR activity of both LmoCas9 and SpCas9. Acr proteins can be used to control...desired edit can increase efficiency. Synchronizing cells or modulating the cell cycle can help as HDR .... CRISPR imaging can visualize chromatin and RNA dynamics in live cells. Labeling can be done by fusing...capabilities , as it can process multiple gRNAs under a single promoter, and can cleave ssDNA non-specifically...and quantification and can be used on lateral flow strips. Other Cas enzymes can be added to SHERLOCKv2...shredder” system that can cause large genomic deletions. The boundaries of these deletions can be defined, either... an enzymatically inactive form of Cas9; can bind, but cannot cleave DNA DSB D ouble- S trand B reak, ...
  7. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...gamma-retroviruses can only infect mitotically active cell types. This means that lentiviruses can infect a greater...proto-oncogenes. This can lead to activation of the oncogene and possible development of cancer. Pseudotyping ...continue their lifecycle and replicate. This feature can be harnessed for stable delivery of various genes...been engineered into gamma-retroviral vectors that can be safely used in laboratory settings. These engineered..., or shRNA of interest into the transfer plasmid can be done with most standard cloning methods, including...Production Using HEK293T Gamma-retroviral vectors can be packaged directly in human embryonic kidney 293T...different envelopes to modify tropism, as the env gene can easily be changed. The HEK293T cells will produce...
  8. Adenovirus Guide

    Type
    Guide
    ... Adenoviridae family and can infect dividing and non-dividing cells. AdVs can cause a range of mild diseases...system recognizes adenoviral elements rapidly and can mount a strong immune response quickly, which has... of interest. The size of the genetic cargo that can be included between the ITRs is limited by the physical..., or shRNA of interest into the transfer plasmid can be done with most standard cloning methods, including...transgene into the adenoviral backbone plasmid. One can also co-transform standard BJ5183 with supercoiled... with the linearized construct, and rAdV vectors can be harvested about 7–10 days later. Vogelstein designed... transgene of interest. With these plasmids, one can track the infection of HEK293/HER911 cells throughout...
  9. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...genome can have on transgene expression and the expression of neighboring genes. rAAV episomes can develop... plasmids can be used to create viral vectors capable of infecting target cells, but cannot produce any...genus, included within the Parvoviridae family. They can infect dividing and non-dividing cells; however, ... of interest. The size of the genetic cargo that can be included between the ITRs is limited by the physical..., or shRNA of interest into the transfer plasmid can be done with most standard cloning methods, including...integration is mediated by proteins Rep78 and Rep68, which can bind to specific Rep-binding elements present in ...processed into a double-stranded circular episome which can concatemerize, producing high molecular weight structures...
  10. Guide to Using Pooled Libraries

    Type
    Guide
    ... use a pooled library; and what types of screens can be done with a pooled library....plasmids with unique, semi-random sequences that can be used for applications like lineage tracing or ...expressing multiple genes at once. A well-designed screen can help you begin to understand what genes are important...population of plasmids in a single tube. Pooled libraries can be small, if they are designed to cover only a subset...the best results. For some libraries, plasmid DNA can be delivered directly to the cells of interest. With...positives or false negatives in later experiments, and can also negatively affect data reproducibility. Figure...biological replicate for that plasmid. Library screens can be divided into two types: positive screens and negative...
  11. Chemogenetics Guide

    Type
    Guide
    ...channels (LGICs) that can either excite or inhibit neuronal signaling. PSAM-based LGICs can be classified by...specific cell populations (most often neurons) that can be turned on or off by the application of a small...neurotransmission. This, and the fact that GPCR ligands can have high potency, which is potentially important...There are several different types of DREADDs that can be broadly classified by the signaling protein that...clozapine and other clozapine metabolites and these can have off-target effects. More recent studies have.... Scientists have shown that both KORD and hM3Dq can be used in the same organism to allow bidirectional...in our Optogenetics Guide . Figure 4: Luminopsins can be activated through chemogenetic or optogenetic ...
  12. Modular Cloning Guide

    Type
    Guide
    ...site, multiple modules with complementary overhangs can be digested and ligated in a single step resulting...resulting in a 4-bp fusion site in between. MoClo can be used for a variety of applications—like building synthetic...sets of cloning vectors (Level 0, 1, and 2) which can be used in successive assembly steps. Before beginning...as in Figure 1). Next, up to six Level 1 modules can be assembled into a Level 2 vector, forming a functional...Dueber 96 standardized and characterized parts that can be used for bottom-up hierarchical assembly of single...Toolkit Yeast Expression Paul Young 34 plasmids that can be used in conjunction with the MoClo Yeast Toolkit...genome integration in fission yeast S. pombe . These can be used with plasmids from the MoClo-YTK . MoClo ...
  13. Promoters

    Type
    Guide
    ...activator can bind), and the structural genes that are transcribed together. Operon regulation can be either...in the absense of lac repressor (lacI or lacIq). Can be induced by IPTG or lactose araBAD Inducible by...in the absense of lac repressor (lacI or lacIq). Can be induced by IPTG or lactose Eukaryotic Promoters...explain how many physically distant DNA sequences can affect transcription of a given gene. There are three...constitutive or inducible expression. Many other promoters can drive expression in specific cells or under different...
  14. Optogenetics Guide

    Type
    Guide
    ...Microbial opsins, such as those described below, can be targeted and expressed in specific subsets of ... neuronal activation/inactivation periods. These can range from msec with hChR2 to "long lasting" (seconds... in the figure above, this light induced binding can be utilized to control the localization of a protein...interest. A genetically-encoded nuclear Cry2 fusion can thus direct a CIB1-protein fusion to the nucleus ...half to CIB1, light-stimulated heterodimerization can reconstitute the protein. This principle has also...applications described above, both LOV and Dronpa can control activity of a fused protein through allosteric....1808 PMID: 22179551 Wietek, J., Beltramo, R., Scanziani, M., Hegemann, P., Oertner, T. G., & Wiegert, ...
  15. Science Guides

    Type
    Guide
    ...use antibodies conjugated to a signaling molecule, can be used to capture, quantify, and identify proteins...
  16. Sequencing Primers

    Type
    Guide
    ...software to see additional annotated primers. You can also use Addgene’s protocol to design your own primers...
Showing: 1 - 16 of 16 results