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We narrowed to 895 results for: Ide

Showing: 301 - 320 of 895 results

  1. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...want to reconsider your approach, and try a different cloning strategy. Once you've identified potential...plasmid and all the steps you will have to take. Decide which technique you are going to adopt (i.e. restriction...digestion on an agarose gel. If the digestion does not provide the fragments of the size you expect, check the...region of the primer to 40-60 bp. It is always a good idea to perform primer optimization, especially if you...that, they can be open source (i.e. primer3) or provided by the major companies selling PCR reagents. Nowadays...Now that you've read our plasmid troubleshooting guide, you hopefully have some new strategies to tackle... the lab he develops new assays and processes. Outside the lab, he enjoy the outdoor in all the possible...

  2. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...capacity of stem cells. Yet, very few researchers consider the sex of their cells when doing experiments—...between male and female cells. The result of not considering the sex of the stem cells was that their male... in Norway, the team realized they should also consider the sex of the stem cells. In this case, they ... many researchers I’ve talked to who’ve never considered the sex of their cells. For one thing, cells ...hard to determine. To remedy this, Shah et al. provide a table indicating the sex of the most commonly... there are numerous methodological articles to consider—we have loaded those we are aware of in our checklist...

  3. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...just isn’t always ideal. But the increased flexibility helps even when it’s not an ideal site: moving several... that you hope is present next to the "perfect" guide sequence for your genome engineering experiment....editing landscape. The need for PAM-less editing   Guides for CRISPR are typically selected by the ‘quality...sequence, that of SpCas9, is NGG. Assuming all nucleotides are represented equally, that’s only a 1/16 chance...flexibility in gRNA selection and subsequent editing. Identifying PAM-free and -flexible Cas enzymes   The solution...more pan-genome PAM flexibility, you’ll want to consider one of several Cas9s that recognize NG PAMs. The...sought to address the PAM problem through structure-guided engineering of the PAM-interacting domain of Cas9...

  4. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    Published
    May 22, 2018, 1:38 p.m.
    ... Time project provides a framework for these undergraduate research questions. The idea is straightforward...were generating huge amounts of data that could provide the basis for student projects in introductory .... Genome projects use computational methods to identify genes by their similarities to genes in other ...studies generally leave questions about gene function wide open. What if two seemingly similar proteins have...phenotype is corrected, students can be relatively confident that the gene from the test species has a similar...Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...about Synthetic Biology with BioBuilder Use Addgene Videos to Learn about Basic Lab Techniques High School...

  5. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...trials involve identifying a therapeutic gene, finding a means to deliver it and identifying a suitable route...technologies have allowed scientists to quickly identify underlying genetic causes of some human conditions...example, it was possible to transduce the T-cells outside the body and thereby correct the mutations using...productive infection. AAV vectors are engineered to provide additional safety benefits: they lack all viral...immunity. AAV vectors are also capable of infecting a wide range of cell types including but not limited to...through direct evolution, capsid shuffling, and peptide displays. These capsids will be selected both for...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

  6. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ... the last year for diagnosing COVID-19, aside from the widely used qPCR and antigen tests.  Labs including...Konermann S, Hsu PD (2021) Genome-wide, bidirectional CRISPR screens identify mucins as critical host factors...spike protein was developed based on 100 structure-guided spike designs and has improved protein yields and...During the 2003 SARS outbreak, Hyeryun Choe’s lab identified that ACE2 is the functional receptor for SARS-CoV..., and more. Recently, scientists used the genome-wide CRISPR libraries, Brunello (knock out) and Calabrese...expressing SARS-CoV-2 open reading frames and identified interactions of these proteins with human proteins...Months later, we partnered with Ginkgo Bioworks to provide synthesized SARS-CoV-2 expression plasmids available...

  7. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...genome-wide library deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... can both process crRNA and carry out the crRNA-guided DNA cutting, unlike other well-characterized Cas...to delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit...researchers sift through thousands of cells to identify cells exhibiting properties of interest. However...of Yellow Fluorescent Protein (YFP). The team identified a new YFP variant, mGold. mGold was found to ...reduction in performance compared to larger genome-wide libraries. Find the library here New from the...

  8. How to Prepare for an Industry Interview

    Type
    Blog Post
    Published
    July 9, 2020, 1:15 p.m.
    ... your video call.  Expect technical difficulties. To try and circumvent this, test out the video conferencing...development, and company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer... homework before the interview. If you are not provided with a list of who will be interviewing you, ask... your audience. Connect your prior work or a new idea to the company’s area of expertise. This has potential...difficulties. Have your presentation on a flash drive and consider bringing your own laptop as a backup, if the company...the interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully...after completing each section of your talk. Some video conferencing software allows participants to “raise...

  9. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ...proteins had to be re-engineered for each gene, making wide-scale gene activation seem next to impossible. The... systems only require changing the programmable guide RNA. Gene activation by dCas9, also referred to ...specific sequences, CRISPR/Cas systems rely on a guide RNA complementary to the sequence of interest. Upon...VP64, a strong transcriptional activation domain. Guided by dCas9, VP64 recruits transcriptional machinery...of VP64 per each dCas9, SunTag uses a repeating peptide array to fused with multiple copies of VP64. By...two-component system dependent on gRNA design (SAM) or peptide design (SunTag). This streamlines its delivery,..., Crawford GE, Reddy TE, Gersbach CA (2013) RNA-guided gene activation by CRISPR-Cas9–based transcription...

  10. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    Published
    Sept. 27, 2017, 1:23 p.m.
    ...example at an anniversary or for Girls’ Day, offer guided tours to show people where the lab magic is happening...share your knowledge at local schools. Or work as a guide in local museums, zoos or botanical gardens. You...Addgene are open to guest contributions. If you decide to start your own blog, check out these two excellent...record a podcast or put some effort into making a video? Use your unique talent to communicate science. ... Guardian, BBC Breakfast or the Londonist. BBC provides a peak into their day-to-day business with popular...Co-Ordinating Centre for Public Engagement (NCCPE) provides on-site courses in public engagement for institutions...media skills, writing about your research and residential communication. If science journalism is the path...

  11. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    Published
    March 15, 2016, 2:30 p.m.
    ...lentiviruses in parallel, this approach should be considered. Similar to the p24 assay, titering via measurement...When using qPCR based methods for viral titration consider using a reverse transcriptase mutant in a separate...carryover. Functional titer Functional titer is considered a more accurate method to quantify virus as it...procedure can take several months but should be considered in labs that will routinely be producing virus...virus or require precise titers. Additional considerations for titering lentivirus Volume and transduction...batch of virus for long term use, you may want to consider using an aliquot that has been subjected to a ... labs that routinely produce virus may want to consider the more accurate proviral integration assay. ...

  12. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    Published
    Jan. 21, 2015, 1:30 p.m.
    ...person is able to communicate their ideas confidently while also considering the needs of other people. Some...Bramson, author “Coping with Difficult People,” identified seven types of difficult people: Hostile-aggressives...postdoctoral fellows are hesitant about expressing their ideas, especially if their supervisors are difficult people...frustration or anger, because your discussion will get side-tracked, and you might create even more conflicts...Your supervisor might not agree with all of your ideas, but he or she probably has reasons for it. Do not... strategies will help you to communicate with confidence earn respect from your supervisor. Whenever possible... resolve most conflicts if you communicate your ideas assertively, listen to his or her viewpoint, and...

  13. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    Published
    Oct. 22, 2014, 1:57 p.m.
    ... asking the panelists not to use any slides for the event. Slides are for presentations, not discussions... the prep calls, but often the prep calls provide good ideas for interesting questions. Be ready to encourage... panelist can contribute. The moderator should identify the funny, provocative or interesting stories ...the actual event, based on these interviews. The idea is to ask questions in a directed way to bring out...discussions. I will rarely work with a panel if slides are allowed. Don't put your panelists behind a table. ...it when the moderator sits in the middle, but the side can work. Only put out enough seats for the expected...more great tips on running panel discussions see Guidelines for Great Panels by Scott Kirsner and How to ...

  14. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ...instead of recognizing nucleotide triplets, the components recognize individual nucleotides. This imparts TALENs...movie Ethan Hawke’s character revolts against class divide that is created by genetics.    Zinc finger nucleases...repair template.  Each zinc finger recognizes a nucleotide triplet and when several ZFNs are strung together...using a protein to recognize DNA sequences is not ideal. For the early generation gene editors, new amino... protein-based DNA-binding domain with a single-guide RNA sequence.  The Cas proteins, just like its predecessor...prospects of curing diseases caused due to single nucleotide variations in the genome.    Due to the rise ...reverse transcriptase. Using an elongated template + guide RNA called pegRNA (prime editing RNA), the prime...

  15. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...polymerase, tRNAs, ribonucleotides and amino acids. Cell-free expression systems are ideal for fast assembly... can then be translated at the ribosome into polypeptide chains that make up the primary structure of ...modifications including protein folding, formation of disulfide bridges, glycosylation and acetylation to create...Mammalian expression systems Mammalian cells are an ideal system for the expression of mammalian proteins ...of the T7 lac promoter and induced by lactose, provides the highest level of protein expression. Despite... and patience. Plant expression systems Plants provide a cheap and low-tech means of mass expression of...recombinant proteins with a naturally secreted plant peptide allows for easier access and purification of a ...

  16. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ... human gut flora. Guides targeted essential and non-essential genes with two guides per gene. The library...expressing guides that target essential genes were less represented while bacteria containing guides targeting... Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined suite of tools for using CRISPRi...Higher transfer rates are better suited for genome-wide CRISPRi library screening experiments where maintaining...biology of bacteria, Mobile-CRISPRi can be used to identify antibiotic resistance genes. Genes that are targets... modular nature allows for the customization of guides, promoters, selection markers, and different versions... Find bacterial CRISPR plasmids Read our CRISPR guide ...

  17. Transferable Skills: Negotiation

    Type
    Blog Post
    Published
    March 3, 2022, 2 p.m.
    ...need some help. Today in our transferable skills guide: Negotiation. As a grad student, I thought that...perspective of all persons involved and be used as a guide to point you toward a creative solution where everyone...specific details about the individuals, each example provides a scenario where both people get something valuable...example shows, the ultimate resolution can involve ideas not related to the actual data collection task but... relationships with your colleagues within and outside of your organization. In my non-academic roles,...investing in me is also beneficial to them? I have an idea on how to improve my current workflow but I need...negotiation  Skills Snapshot: Negotiations When Cultures Collide: Managing Successfully Across Cultures by Richard...

  18. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ...procedure The procedure (illustrated in the YouTube video below) is simple: After obtaining antibiotic-resistant...form a firm monolayer. The cell clones - easily identified from the plate circling- are then individually...plug by rapid pipetting, as shown in the YouTube video. Before returning the plate to the incubator, additional... a part of the body very sensitive to heat- the inside of the wrist or the cheek. (Parents of infants ... well separated. For this, you need to have some idea of the rate of growth of your specific cell line...cells we sometimes use TPP peelaway T-75 flasks to provide a greater surface area (the amount of agarose/DMEM...transfected clones. The technique is applicable to a wide variety of cell lines, from HEK to CHO cells to ...

  19. Making Career Development an Integral Part of the Science Training System

    Type
    Blog Post
    Published
    Feb. 25, 2021, 2:15 p.m.
    ... of their undergraduate experiences, they were provided with ample resources and a positive perspective...In 2018, the ASBMB and Burroughs Wellcome Fund provided seed funding to take this initiative to the next...just released report “Enhancing Dissemination of Evidence-Based Models for STEM PhD Career Development; ... for pd|hub which include building capacity to identify the most effective practices and disseminating...training approaches in a way that can be implemented widely in the community.  Talk alone is not going to drive...development and implementation of collections of evidence-based practices. These concrete programs and practices...PhD professional development are intended to be widely disseminated by “train the trainer” programs.  ...

  20. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...fluorescent dye or HRP via maleimide-labeling. Maleimide-labeling occurs when a maleimide group forms a covalent...found in cysteine. Maleimide-conjugated HRP can be found here and several maleimide-conjugated dyes are...reporter enzyme. For multicolor staining, careful consideration must be used when selecting which antibodies...only one primary antibody. This process can be avoided direct antibody fluorophore conjugates are used..., you can learn more about how nanobodies are identified in this review. If you just want to use the nanobodies...nanobodies are outlined below; no alpaca required. Decide Which Nanobody To Use: The table below summarizes...
Showing: 301 - 320 of 895 results