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We narrowed to 74 results for: tes/1000

Showing: 21 - 40 of 74 results

  1. Promoters

    Type
    Guide
    ...interest is expressed. Summary Promoters are about 100-1000 base pairs long and are adjacent and typically upstream...the other hand, the process in eukaryotes is much more complex. Eukaryotes require a minimum of seven transcription...actually begins. Only eukaryotes and archaea, however, contain this TATA box. Most prokaryotes contain a sequence...are common in prokayotes, specifically bacteria, but have also been discovered in eukaryotes. Operons consist...binding is very different in bacteria compared to eukaryotes. In bacteria, the core RNA polymerase requires...Transcription is tightly controlled in both bacteria and eukaryotes. Promoters are controlled by various DNA regulatory...promoter also contains transcription factor binding sites, but mostly contains regulatory elements. Eukaryotic...

  2. Antibody Guide

    Type
    Collection
    ...sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process may...response has developed, the antibodies are collected, tested against the antigen, packaged, and sold. This is...immortalized B cells). The genetic reorganization that creates variety in the antibody allowing for their extreme...class therefore has a valency, or number of binding sites available per antibody unit, which is typically ...multiplied by two, as each antibody has two binding sites. Isotype Description Configuration and Valency Subclasses...Monomer with a valency of 2 IgE Protects against parasites and is responsible for driving allergic reactions...in the lab. Check for and record any expiration dates. Buffers Buffers are a concern for conjugating and...

  3. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...produces point mutations at a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate..., et al. F1000Res. PubMed PMID: 27830062 Toxoplasma CRISPR knockout pooled library Parasites from the...availability of enhancer sequences in different chromatin states. Alexander Stark and colleagues at the Research...self-transcribing active regulatory region sequencing) that tests enhancer sequences outside of their endogenous chromatin...Synthetic and basic biologists alike often need to test multiple expression systems before they achieve ...bacteria, the process of cloning, combining, and testing these different parts can be quite arduous. The... lab generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...

  4. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...sector throughout their careers (McDowell et al, F1000Research, 2014; Dolan, F100Research, 2016). These events...of the Federal Labor Standards Act (FLSA) which mandates that all workers across the country either be ...specifically chosen to better define productive routes for advocacy efforts and help participants develop...Fisher Scientific Developing effective science advocates is an important way to push the balance from challenge...Research: a perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...from the Future of Research Chicago Symposium F1000Research, 2016. Kyle T. Dolan, Joseph F. Pierre and Erin... advocates...

  5. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...new target sites. Created in BioRender.com.   Class II TEs: DNA transposons Class II TEs are also ...of transposition: Class I TEs, also called retrotransposons, and Class II TEs, also called DNA transposons.... Class I Transposable elements (TEs): Retrotransposons  Class I TEs are also known as retrotransposons...the TE while also increasing the size of their host’s genome.   There are two types of Class 1 TEs: those...DNA sequences or target sites. Integration of most transposable elements (TEs) results in the duplication...into new target sites. This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode...encode a transposase enzyme.  Class I TEs are considered replicative since every time they jump they make...

  6. Molecular Biology Reference

    Type
    Guide
    ...fluorescent signal. Sanger can sequence approximately 500-1000 bases downstream of the known primer region with...of DNA, or templates, are spatially separated from each other. These attached DNA templates are then amplified...Alternative start codons include CUG in eukaryotes and GUG in prokaryotes. Common Epitope Tags Tag Amino Acid...DNA. Although they can be found in archaea and eukaryotes, they play the most significant biological role...fragment with a plasmid or vector DNA backbone generates a recombinant DNA molecule, which can be used ...of DNA which contains several restriction enzyme sites, enabling easy insertion of DNA by restriction enzymes...the promoter. As a general rule, the restriction sites in the MCS are unique and not located elsewhere ...

  7. Dos & Don'ts When Publishing a Scientific Manuscript

    Type
    Blog Post
    ...Maaike Pols, PhD, a Developmental Editor at F1000Research.  With many changes taking place in the world...your Addgene plasmid #), and other reagents. F1000Research, among many other journals, has joined a pilot...results more valuable. Some journals, including F1000Research, also ask for the raw data behind the figures...several rounds of revision before acceptance. F1000Research uses a novel form of peer review that occurs...sure your manuscript fits the house style and prerequisites of the journal. Ask colleagues (preferably native... Maaike Pols is a Developmental Editor for F1000Research, responsible for assessing the scientific content...

  8. Antibody Guide

    Type
    Guide
    ...sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process may...response has developed, the antibodies are collected, tested against the antigen, packaged, and sold. This is...immortalized B cells). The genetic reorganization that creates variety in the antibody allowing for their extreme...class therefore has a valency, or number of binding sites available per antibody unit, which is typically ...multiplied by two, as each antibody has two binding sites. Isotype Description Configuration and Valency Subclasses...Monomer with a valency of 2 IgE Protects against parasites and is responsible for driving allergic reactions...in the lab. Check for and record any expiration dates. Buffers Buffers are a concern for conjugating and...

  9. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

    Type
    Blog Post
    ... organization issues – F1000 just announced the launch of their F1000Workspace.  Give your favorite searching...your search and sorting the results (relevance, dates etc). The links for the titles can generally take...you provide, Google Scholar will send you daily updates with relevant literature. Organizing the literature...margins. If you prefer, printing and hand-writing notes is still possible, but not feasible for every paper... University. She is a fan of photosynthesis and writes about autotrophs and biochemistry on her blog. ...Maaike Pols, PhD, a Developmental Editor at F1000Research  ...

  10. Lentivirus Production

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -20 °C...PEI such that the ratio of μg DNA:μg PEI is 1:3 (1000 μL total per 10 cm dish). Using transfer plasmid...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ... plate in DMEM Complete in 10 cm tissue culture plates. Incubate the cells at 37 °C, 5% CO 2 for ~20 h...the table below for a possible range of ratios to test: Ratio of DNA:PEI μg of DNA μL of 1 mg/mL PEI 1:...Centrifuge the viral supernatant at 2100 rcf for 5 minutes to pellet any packaging cells that were collected...

  11. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...reasons, it is essential to test the selectivity of your antibody. But testing is not one size fits all....important to test for the protocol you intend to use in your experiment. An antibody that demonstrates selectivity...best to determine this by testing them. For intracellular targets, we usually test 3 different fix/perms ...and flow cytometry. F1000Research, 13, 817. https://doi.org/10.12688/f1000research.154034.1 Laflamme, ...synaptotagmin 1 page. The data produced is available on the F1000 gateway and Zenodo community.  Knockdown approach... Correctly interpreting your data therefore necessitates careful design of RNAi sequences, confirmation...like a blood sample containing many different leukocytes. Where data is available from antibody-independent...

  12. AAV Production in HEK293 Cells

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Hemocytometer or cell ..., Nalgene, 565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration Unit, Nalgene 167...solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -80 °C... deionized water and adjust to a final volume of 1000 mL. Stir at RT until fully dissolved. Pro-Tip This...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ...inactivated in the lab by heating to 56 °C for 30 minutes. 0.45 μm polyethersulfone (PES) filter system, ...and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently tap the sides of the...

  13. Bacterial Expression Systems

    Type
    Collection
    ...Consortium Modular Assembly Kits (Addgene #1000000005, 1000000010) contain plasmids and an engineered strain...being used for their distinct features. Addgene distributes many plasmids that can help researchers plan ...western blots, or removed using protease cleavage sites. Other protein tags and signal peptides are used... reduces bacterial protein contamination and facilitates extraction. Browse our most popular plasmids ...protein purification. Commonly used tags, cleavage sites, and signal peptides include: Epitope tags: 6xHis...Solubility tags: MBP, SUMO, TrxA, Mocr, NusA Cleavage sites: TEV protease, factor Xa, enterokinase, thrombin...localization to the periplasm. The pCri System (Addgene #1000000058) plasmids can be utilized for heterologous cytoplasmic...

  14. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    ...you want to have 100x-1000x coverage of your library: in other words, 100-1000x more reads than number...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds,...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...solid media. The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library...using both pre- and post-amplification DNA and generates PCR sequencing products from both. These products...

  15. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen.1000683 Bétermier M, Bertrand P...finishes repair of most types of breaks in tens of minutes – an order of magnitude faster than HDR. NHEJ is...paired end complex The paired end complex then ligates compatible DNA ends together, thus repairing the...Aprataxin, Tdp2) that function to make ends better substrates for ligation. Although we do not describe these...vastly reduces breaks and mutations at off-target sites. Christina Mork contributed to the update of this...

  16. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system facilitates the simultaneous... Viruses are intracellular parasites and natural vehicles for genetic information. Therefore they make...Addgene: Follow our viral vector blog posts for updates on the most recent deposits, protocols, and tips...Plasmid Kit Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of...

  17. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...mL conical collection tube. Centrifuge column at 1000 x g for 2 min to remove storage solution. Place ...container. Add 5 mL PBS to the column. Centrifuge at 1000 x g for 2 min to remove PBS. Repeat Steps 30–31 ...resin bed to fully absorb the sample. Centrifuge at 1000 x g for 2 min to collect the sample. Discard column...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...tubes, VWR 76289-498 Before Starting Wipe down all pipettes and equipment with 10% bleach prior to use. Warm... Adjust pH to 7.0 Autoclave or filter sterilize 1000X protease inhibitor cocktail 25 mg leupeptin 50 mg...Spectrophotometer using the A280 IgG setting. Combine all eluates with measurable protein. Determine the protein ...

  18. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller...96-well plate, seed one of the corner wells with ~1000 cells. This will help you focus the microscope later... incubating the cells. Pro-Tip Use the well with 1000 cells to focus the microscope, then you can easily...PES filter or centrifugation at >500 x g for 5 minutes. Do not use the medium if the cells are overly ...

  19. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 O’Brien, R. E., Bravo, J...unwinding of dsDNA occur simultaneously, which creates a loop in the target strand that is cut occasionally...

  20. Typing CRISPR Systems

    Type
    Blog Post
    ...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on ... clustering and discovery algorithms find new candidates for CRISPR proteins. A type VII is already on...
Showing: 21 - 40 of 74 results