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We narrowed to 798 results for: omp

Showing: 121 - 140 of 798 results

  1. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...transformation First, use enough DNA and competent bacteria to ensure all components of the library are represented...represented. Typically, Addgene uses electrocompetent Stbl4 cells when performing amplifications, and we use electroporation...may be amplified preferentially, skewing the composition of the final recovered DNA. This bias can be ...need to count how many times each of the library components is found in the read list. If you also sequenced...sequenced the pre-amplification DNA, you’ll compare the numbers before and after. However, because there are...give you a bell curve of the abundance of each component of your library. You should look for two things...making sure few/no plasmids have dropped out, and comparing pre- and post-amplification sequencing results...

  2. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...reduced transformation and transfection efficiency compared to the monomeric version of the plasmid (Maucksch...multimers have additional origins of replication compared to monomers, they are also able to replicate at...bacterial cells with multimers grow more slowly compared to those containing the monomeric version of the...and more prone to being lost in bacterial cells compared to monomers, although this can be prevented via...cannot be used to detect plasmid multimers. Some companies provide NGS whole plasmid sequencing services ...frequency of multimerization, and in some cases completely prevent the formation of plasmid multimers (Bedbrook...produced the highest yield of monomeric plasmid compared to DH5α. However, more research is needed to determine...

  3. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...larger datasets and use computer based analysis techniques, an important component of reproducibility is... for preclinical studies, as reported by drug companies attempting to apply the findings from these studies...research. I’ve talked to many researchers who have complaints about the unreliability of antibodies. IPI is...technologies to your own research needs. Other companies like NEB and ATCC can provide you with additional...sifting through paper lab notebooks). Once you’ve completed your experiment, you’ll need to analyze your results...can understand your analyses and repeat them. Companies like Code Ocean allow researchers to share code...submit designs and transparently reveal them upon completing an experiment. There’s not necessarily in-principle...

  4. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    Published
    Feb. 28, 2023, 2:15 p.m.
    ...achievement, the complexity of the device increases, as does the price. With this increased complexity comes the...consumer-grade components. In this field, often referred to as 'frugal science', existing components are adapted...are constantly being developed which combine computational processing with biological protocols and revolutionary... rarely provide the construction plans or CAD (computer aided design) drawings of their devices, which...around the use of commercially available tools and components. Rapid prototyping techniques such as 3D printing...and can allow for innovative organizations and companies to provide problem-solving platforms.     ...

  5. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...previous blog post about how they started their company here. The first transgenic plant was engineered...cannot build what we do not understand, and the complicated and unique metabolic processes of plants are ... Our color changing flower involves two basic components: the pigments (anthocyanins), and the plant circadian...also identified and characterized mechanisms and components of the circadian rhythm in petunias. Because ...before us, it’s possible for us to combine these components to develop flowers that change color. Without...Without this basic research – this comprehensive understanding of the genetic pieces underpinning these mechanisms...plant synthetic biology. There are already two comprehensive plant cloning kits available in the repository...

  6. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ... as these immune complexes are smaller than a primary:secondaries:conjugates complex. If you are trying...and therefore each target protein. This can be accomplished by using a panel of primary antibodies conjugated...tissues and cell lines; orthogonal strategies that compare antibody assay results with those of antibody-independent...transcriptomics; independent antibody strategies that compare the staining patterns of multiple antibodies against...that express a tagged version of the antigen and compare staining of the antibody to that of the anti-tag... strategies that isolate an antibody’s immune complexes and confirm interacting partners by mass spectrometry... guaranteed to work. Antibody-based assays are complex and antibodies may perform differently in different...

  7. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    Published
    May 24, 2018, 1:20 p.m.
    ...worm amputations. They also created an online companion resource called “cutting class” which provides...Digoxigenin-labelled Uracil) that will bind to complementary mRNA within an embryo or tissue. Once the riboprobes...specific antibodies that detect the mRNA-riboprobe complex are added to the tissue. The last step involves...cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue. Students...organism of interest, use the RNA to create a complementary DNA (cDNA), and then use PCR and molecular cloning...performing these experiments have a good baseline for comparison. Performing WISH using these plasmids can result...one strand of the siRNA and are directed by complementarity to the remaining single stranded siRNA to bind...

  8. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...RNA is a structural and functional component of the ribosome complex and Dr. Steitz quipped that RNA in...development zone so many science and technology companies and start-ups choose to locate there. I made time...was the outdoor market where one can shop for components of traditional Chinese medicine. There were hundreds...Lowe lab have allowed them to begin understanding complex regulation of and by tRNAs and tRNA-derived RNAs...replication with no proteins). The chemistry is complex and this work requires an interdisciplinary team... they do it. Of course, No conference would be complete without CRISPR Michael Terns from of the University...in the ribosome is in a complicated conformation “not like DNA which has a boring structure.” That certainly...

  9. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...alternative PAM sequences are longer and often more complex than NGG, resulting in lower overall genome frequency...Characterizing xCas9 in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells...PAMs averaged 56-91% transcriptional activation compared to the NGG PAM, confirming the flexibility of ...with improved editing at an NGG PAM (69 vs 48%) compared to SpCas9-containing ABE7.10, as well as improvements...NGS) are shown as percentages, with fold changes compared to SpCas9 in parentheses. The development of ...that selecting for one beneficial trait, PAM compatibility, also produced an enzyme with improved on-target...

  10. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...when studying them since their complete knockdown is lethal. Incomplete or titratable knockdown, however...generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission... Top: A Tn7 transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes...ability to synthesize diaminopimelic acid (DAP), a component of the cell wall. Once in the recipient cell, ...Bottom: An ICE element containing the CRISPRi components is transferred to recipient bacteria by bi-parental...was variable. They measured transfer rates by comparing the number of colonies that grew on a selective... gRNA were more sensitive to trimethoprim when compared to control gRNA treated or untreated bacteria....

  11. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ... experiments are also generally comparable so the best way to compare costs is to look at the prices of...In today’s post, we will focus on a high-level comparison of popular cloning methodologies, with a special...least 50 cloning reactions. The ligase cost is comparable to that of the restriction enzymes and overall... in a price of a few dollars per reaction. By comparison, Gibson assembly can cost over $15.00 per reaction...more streamlined, because they will come with comprehensive and tested protocols, as well as premixed and..., like Gibson or SLIC, since one can pool the components and define their final order through sequence...Golden Gate assembly allows one to simultaneously complete an ordered set of scarless ligations following...

  12. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...relatively easily synthesized, must be produced using complex mixtures derived from cells or using live cells...systems do require demanding cell culture conditions compared to other systems. Insect expression systems Insect...Insect cells can also be used to produce complex eukaryotic proteins with the correct post-translational.... These systems enable high expression of very complex, glycosylated proteins that cannot be produced ... expression systems, including the majority of complex post-translational modifications. Extraction and... as plant tissues themselves are biochemically complex. To circumvent these issues, scientists have taken...mammalian cells, and are often capable of modifying complex proteins unlike bacterial systems. Yeast cells,...

  13. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...realized how perfectly flat the complex is on this surface”….where the complex is docked on the membrane of...using 3D printing technology. The tagline for our company is “…where molecules become real”.  We sometimes... the real thing. Nevertheless, the models are compelling, and I encourage you to hold one. Why do we ... They then start talking to themselves as they compare the features displayed on the model with their ... close to its reported cleavage site on the complementary DNA strand. And this is why we love working ... that was not “poised for cleavage”. Jacob computationally added the non-template strand to the Jinek ...double-stranded DNA, recognize a PAM sequence, allow the complementary base pairing between the guide RNA and the target...

  14. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ... millions of people die from cancer-related complications each year. Despite these grim statistics, new...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and tumor suppressors...cancer research has focused on picking apart the complex cellular pathways involved in the development of...for this elusive target. Recent improvements in computer modeling have enabled researchers to begin discovering... prevalent in human cancers, and synthesizing compounds to bind specifically to their mutated residues...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...

  15. Antibodies 101: Designing Your First Flow Panel

    Type
    Blog Post
    Published
    May 28, 2024, 1:45 p.m.
    ...antibody-conjugated labeling. Even though the two compounds are completely different in nature (eGFP being a protein...results in a higher-wavelength emission spectrum compared to APC. This happens due to a process called Fluorescence... red laser excites at 640 nm (Table 1), while comparing the two fluorophore's excitation spectra around...think of brightness levels on your smartphone or computer screen), and you should be able to properly label...the same color panel. In this case, make sure to compare the excitation/emission spectra of your reporter...whether and to what extent they overlap. Figure 4 compares the spectra of enhanced GFP (eGFP) and FITC, the...Make sure to check out our blog post on color compensation to learn more about them. Alright, let's build...

  16. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    Published
    July 27, 2023, 1:15 p.m.
    ...referred to as “complex retroviruses ” (Cullen, 1992) because of their more complicated genomes. Here, ...biology. “Retroviruses” is an umbrella term which encompasses 1.) the gamma-retroviruses — that have been developed...whole family; however, you can use Figure 1 to compare the genomes of these two family members.   ...beginning to realize that the situation is already complex enough that a deep understanding of retroviral ...Because it is made at the same time as the other components of the vector particle, some of the RT protein...In both cases, the lipid membrane is the only component that is not dependent on viral genes. It is derived...Mechanism of action of regulatory proteins encoded by complex retroviruses. Microbiological Reviews, 56(3), 375...

  17. Summer SciComm Series: Modes of Communication

    Type
    Blog Post
    Published
    July 1, 2022, 1:15 p.m.
    ... if you can begin simply and build up to more complexity if needed. And since we live in the digital age... biology and ecology can make for beautifully compelling photos and videos, while molecular, micro, and.... Can you convey enough information to tell a compelling story in an image or do you need a full exhibit...fair, or a semester-long after school program? Comparing your desired outcome to the limitations and potential...expertise needed can increase exponentially with complexity. Podcasts Medium Easy for audience to engage... needed. Traditional media formats are quite competitive while digital can be oversaturated. Transition...Photography Medium (high for microscopy) Visually compelling. Easy to convey a large number of details without...

  18. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...of the vector components are HIV-based? A7: No single plasmid contains all the components necessary to ...The components are divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs...decreasing the probability of creating replication-competent lentivirus (RCL) in your target cells.  Q4: How...targeted. Q6: Are transfer plasmids replication competent? What is SIN? A6: Most (if not all) Addgene transfer...LTRs, PPT, RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more ...are: 1) the potential generation of replication competent virus [usually HIV-1]; and 2) the potential for...

  19. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ...have the full picture of the puzzle, the QC team compares it to the plasmid information or depositor-provided...depositor-provided sequences to confirm the correct plasmid components. For more information on this process, see our...small sample sizes, small regions of interest, or complex sequences such as G-C rich areas, secondary structures... NGS technology, due to the high-throughput compatibility. However, there are some instances where we ...thousands or even millions of base pairs. Several companies have worked on this technology, but Oxford Nanopore...sequence an entire plasmid in one read. Runs can be completed in anywhere from under an hour to 72 hours, with...data output. This method is great for resolving complex and repetitive sequences, but can be costly, low-throughput...

  20. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    Published
    June 20, 2024, 1:15 p.m.
    ...the cell must have enough nutrients available to complete the entire round of replication. Initiation is...initiate replication. But pSC101 requires a third component: RepA, a protein encoded by pSC101 itself. RepA... are efficient for the host, as no additional components are necessary. Iterons are not the only tool ...Inc, also encoded by the ColIb-P9 plasmid, is complementary to part of the sequence forming this pseudoknot...restraining further replication.   Figure 3: Comparison of replication regulation mechanisms of stringently...replication. At high copy numbers, RepA proteins form complexes between iterons on different plasmids in a process...propagation by limiting their copy number and compensating for the additional metabolic cost imposed on...
Showing: 121 - 140 of 798 results