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We narrowed to 520 results for: HA tag

Showing: 281 - 300 of 520 results

  1. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...set as 1. Some proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be ...Fluorescence is the emission of light by a substance that has absorbed light. The emitted light is at a longer ...protein Excitation & Emission (ex/em): Each FP has its unique ex/em peak. Therefore, choose FPs that...detect the emission. For example, if your microscope has only two lasers, at 488nm and 561nm, you will not...pHTomato (pH sensors), HyPer (H2O2 sensor), ArcLight (voltage sensor), and iGluSnFr (glutamate sensor). More ...can fold in <10min at 37°C, mCherry takes ~15min, TagRFP ~100min and DsRed ~10hours. Temperature: FPs maturation...

  2. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

    Type
    Blog Post
    ...reprogrammed less efficiently, no such correlation has been found between the age of the patients from whom...potentially promote tumorigenesis in vivo. The phenomenon has been recorded in past gene therapy trials and remains...reprogramming factors To date, the mRNA transfection method has not been used successfully to derive iPSCs from PBMCs...However, reprogramming of PBMCs by episomal vectors has yielded hiPSCs at a low efficiency (0.001-0.03%) ...efficiency: the pCXLE episomal vector toolkit It has been shown previously that suppression of TP53 enhances...hiPSCs than age-matched fibroblasts [3]. These advantages make PBMCs the ideal source of somatic cells ... This is to avoid the chances of insertional mutagenesis, which is the integration of transgenes into ...

  3. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...Recently, Xue Sherry Gao’s lab at Rice University has developed DAP arrays for efficient multiplex base...process and release its own gRNAs from a single array, has been a top choice for multiplex gene knockout or ...proper functions to enable efficient base editing and has yet to be developed for prime editing, both and their..., which is conserved in all living organisms and has previously been used for multiplex Cas9-nuclease ...lengthy promoters, such as U6 or EF1α promoters, has limited the number of gRNAs that can be packaged ...CRISPR tools, without compromising their strategic advantages in gene editing, while keeping the system to ...

  4. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...genome can occur, although at a low frequency which has been estimated to range from 0.01% to 3% depending... well-characterized and extensively studied, and has a broad tissue tropism. Table 1 gives a summary of...across a broad range of cell types in vivo. AAV-DJ has been optimized further for specific cell and tissue... tropism towards specific cell types of interest has been to create and test capsid libraries with random...systemic capsids. A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...naturally cross the BBB. The Viviana Gradinaru lab has developed a number of systemic rAAV vectors that ...preps . AAV retrograde (AAVrg) Directed evolution has also been employed to engineer an rAAV serotype variant...

  5. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended... antibody libraries to generate new sdAbs. There has also been some interesting work done developing transgenic...al., 2021.   The single-domain antibody field has come a long way since the serendipitous discovery...reagents. But for a quick refresher, some of the advantages of sdAbs over conventional antibodies include... simple structure    sdAbs in research These advantages have contributed to sdAbs becoming valuable tools...

  6. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...positions. There has been a lot of engagement with the Athena SWAN Charter and it has really strengthened...their pathogens. As Patron explains it, her work has always been focused on gene transfer, from transgenes...MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and enable plant science...quantitative data for regulatory elements, which has not been available for plants. Addgene: Will the ...strengthened the network of female scientists. Twitter has been really good for that too. I’m more in touch with...s research network here. She talked about the advantages of talking to people and representing yourself...

  7. Adenovirus Guide

    Type
    Guide
    ...immune response has become a disadvantage for the use of rAdV vectors in gene therapy but has, however, proven...response quickly, which has limited their use in therapeutic applications. However, this has also made them the...the human population has been exposed to at least one human AdV serotype and has developed a serotype-... digest patterns to determine that the transgene has been inserted into the adenoviral backbone plasmid...other immune-modulatory molecules. Another strategy has been to use oncolytic rAdV vectors, which are replication-competent... of spontaneous reversion to RCAs, although this has been reduced by providing the E1 gene in trans within... adenovirus (RCA) Mutated adenoviral vector that has the ability to replicate and reproduce within host...

  8. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...tools. The advantages provided by chemogenetics are instead that it is noninvasive and has longer-lasting... disrupting normal brain function. Chemogenetics has become a popular tool in the field as it provides... agonist for muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have...are already approved for clinical use. Varenicline has been further modified to create ultrapotent PSEMS...will promote hyperpolarization. PSAM ligands The advantage of PSAMs lies in their ability to be specifically...

  9. Help Us Name Blugene’s New Virus Friend

    Type
    Blog Post
    ... favorite! Our mascot who has supported us over the past eight years now has a new friend to join them...taking name suggestions using this form or the hashtag #BlugeneVirusFriend on Twitter. As with Blugene...

  10. Lentivirus Production

    Type
    Protocol
    ...10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through a 0.22 μm...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Low serum medium such as Opti-MEM...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...add 55 mL of heat-inactivated FBS and 5 mL of glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below...

  11. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    ...Davis School of Medicine on this grant. Dr. Trimmer has created thousands of high-quality antibodies/nanobodies...mission This is the first NIH grant that Addgene has received. Addgene prides itself on being financially...The timing is ideal as the NIH’s BRAIN Initiative has been funding laboratories, such as Dr. Trimmer’s ...antibodies are plasmid-based and offer numerous advantages, including more secure archiving, unambiguous...AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS...

  12. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    ...never went away. While he has frequent bouts with Impostor Syndrome, Joe has found his people through ... match people within similar fields (ASM Microbe has eight tracks), something we didn’t have enough people...notebook paper, despite the fact that it was gray. He has worked in molecular and microbiology for the last...conference hashtag I had, however, started a Twitter account, and was following the meeting hashtag. I noticed...Favorable conditions - Expansion of networks The last stage is Exponential Growth, named after the growth phase...

  13. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...and insert a linear fragment of DNA (insert) that has been cut by compatible restriction enzymes. An enzyme...one found on the left side of the image below. It has a promoter (blue arrow) followed by the restriction...finished plasmid   Once it looks like your ligation has worked, you will need to pick individual bacterial...purpose. Ideally, once you know that your plasmid has an appropriately sized insert, you should send it...plasmid for more details. Once your complete plasmid has been verified, you’re ready to get experimenting!...gel purification. In gel purification, you use a voltage difference across a gel matrix (usually agarose...positioned toward the cathode (- charge). When the voltage is applied across the gel, the DNA migrates toward...

  14. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ... but inadequate access to reagents and equipment has become a bottleneck. In the last few years, scientists...nucleic acids at the atomolar level. The Zhang lab has adapted natural RNase activity of the Cas13 protein...Gootenberg et al., 2017 and 2018). While the Doudna lab has used Cas12a’s non-specific ssDNA degradation to develop...field (Myhrvold et al., 2018). Now, the Zhang lab has shared a protocol for using SHERLOCK to detect SARS-CoV...at atomolar levels, within one hour. DETECTR thus has the ability to rapidly detect nucleic acids with ...diagnostic applications. The company Mammoth Biosciences has been started based on DETECTR technology with the...Biotechnology. Like the SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing...

  15. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    ... makes software good? Fortunately, that question has been thoroughly answered by experts in human-computer...specialized controls available when needed. SnapGene has been engineered to be easy and enjoyable to use. ...protein sequence match even when the DNA sequence has changed. Identifying plasmid control elements Coding... with these extensive efforts, such an algorithm has limitations. For instance, it could miss a common...beginning, shepherding a new product from the idea stage through all of its post-launch iterations.     Additional...

  16. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...applications. The potential of CRISPR has yet to fully taken advantage of, but exciting new tools like these... Cas11-like protein, and a guide RNA. The enzyme has dual activities: processing pre-CRISPR RNA into mature...kilobase-sized gap in the gene editing field. The AbuGoot lab has now developed PASTE, which can “drag and drop” kilobases...PASTE is more specific and efficient than HITI and has higher purity of insertion than both HITI and HDR...without detectable non-specific activity. Its major advantage is the significantly reduced amount of off-target...

  17. Colony Formation Titering Assay

    Type
    Protocol
    ...mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then there are 75 colonies /1.5 mL x...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Polybrene...L-alanyl-L-glutamine (or stable alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...55 mL of heat inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots...

  18. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    ...peer mentoring circle. This type of program has many advantages and I am seeing a lot of success at universities...makes a good mentoring program. If your organization has a mentoring program, consider tapping into it and... have going on locally. Your university probably has an alumni directory. Alumni who enter their information...confidential topics Broaden your network Take advantage of formal programs A formal mentoring program ...

  19. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...foot shock that has become associated with a visual or auditory cue. The hippocampus has long been known...every other advanced technique in neuroscience it has a bit of a steep learning curve and many opportunities...within your own neuroscience sub-discipline that has published using the technique and ask if they would...sophistication of mouse transgenics. Everything that has been described is intended for the mouse; however...behavioral models. Beyond rodents, optogenetics has even been applied to other species such as non-human primates...the ferrule implants and patch cables. One huge advantage of using rats is their superiority as behavioral...

  20. Editor's Choice, August 2016

    Type
    Blog Post
    ... Our Editor's Choice Blog Posts The Addgene Blog has had quite the up-tick in views over the month of ...recommend taking a look at all of our posts as each one has something great to offer, I just need to get back...optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course... in chronological order here: Site Directed Mutagenesis by PCR Google Forums Round Up: First Impressions...
Showing: 281 - 300 of 520 results