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  1. Celebrate National Book Month with Addgene

    Type
    Blog Post
    ...larger epic of fantasy novels. Set in a world where 1000 years ago the hero lost and the Dark Lord rules ...Napoleonic Wars and against something even more sinister. 1000 pages long. Incredibly imaginative, funny, bittersweet...emotions, and even what therapeutic properties it has. As with other works of Dr. Sacks, Musicophilia includes... pretty biased as the founder, but our book club has become one of my favorite parts of Addgene’s culture...
  2. The Blue Flame Award: Celebrating Addgene's Most Requested Depositors

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    Blog Post
    ...indicates that a plasmid has been requested 20+ times   A red flame means a plasmid has been requested 50+ ... but a select few are particularly special. Over 1000 of our 60,000+ deposited plasmids have been requested... Castro of University of Michigan says, “Addgene has transformed the way in which we share plasmids and...making it easier, cost effective and faster; this has helped accelerate scientific discoveries.” Occasionally...+ times   A blue flame means a plasmid has been requested 100+ times Since we so sincerely appreciate ...have at least one plasmid deposited at Addgene that has been distributed more than 100 times. We hope these...sharing because, as UNC’s Bryan Roth says, Addgene has “literally enabled sharing with 100's of investigators...
  3. The Power Behind NGS Plasmid Validation: seqWell

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    Blog Post
    ...barcodes. Rather than continuing to process 100s to 1000s of samples individually, plexWell™ reduces every...with our plasmid assembly pipeline. To date, this has resulted in high quality NGS-based assemblies for...
  4. The time and cost required to make a plasmid

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    Blog Post
    ...five years, inflation has occurred. Here at Addgene, the consumables cost per stab has increased 50% since... very cool 8,768,560 hours – or almost exactly 1000 calendar years!   *Dammit, man, I’m a scientist, ...make a plasmid. Doench’s lab in the Broad Institute has created hundreds of thousands of plasmids, using ...since 2019, while overall inflation of USD has gone up 19.8% in the same time period. For ease of calculations...per plasmid, Addgene, since its founding in 2004, has saved the scientific community a collective and very...
  5. Antibodies 101: ChIP

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    Blog Post
    ...pieces of roughly equal size, typically between 200-1000 bp. This is to ensure that your downstream analysis...an antibody for the first time, look for one that has been validated in multiple assay types. Remember ...
  6. Tips for Titering Your Lentiviral Preps

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    Blog Post
    ...between a cell that has had one viral particle integrated into its genome and a cell that has had multiple integration...particles. Studies have found that values can be 10-1000 fold higher than for functional assays depending...The day has arrived; you’ve painstakingly cared for your packaging cell line, prepped your DNA, transfected... be significantly higher than the same prep that has been stored in the freezer. If you plan to freeze..., you may want to consider using an aliquot that has been subjected to a freeze/thaw cycle for titering...CRISPR Genome-Wide Screening Learn How Lentivirus Has Been Used for Lineage Tracing Additional Resources...
  7. Visualizing Protein Turnover In Situ

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    Blog Post
    ...proximity ligation." Nature methods 3.12 (2006): 995-1000. PubMed PMID: 17072308.  4. Dieck, Tom, et al. "...Singer’s lab at Albert Einstein College of Medicine has recently published techniques to follow translation...Rojas! Eugenia Rojas, PhD is a neuroscientist who has always been deeply interested in neurobiology.  She...
  8. Which Fluorescent Protein Should I Use?

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    Blog Post
    ...coefficient and quantum yield of the protein, divided by 1000. In many cases the brightness is compared to that...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...Some proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be taken into...
  9. Droplet Digital PCR for AAV Quantitation

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    Blog Post
    ...used to calculate the viral titer: GC/mL = {[(R*C)(1000/V)]*D} R = Reaction volume C = Copies/uL V = Volume...we should first note that quantitative PCR (qPCR) has been a powerful tool for quantifying AAV for some... 2 minutes and the denaturation time to 1 minute has been shown to increase droplet separation (Witte ...
  10. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ... (response elements). Promoters are about 100 to 1000 base pairs long and found upstream of their target...transcript, you will also need to make sure your plasmid has a promoter suited to working in your host organism... but requires SP6 RNA polymerase. SP6 polymerase has a high processivity. When used for in vitro transcription...
  11. Uncovering Ancestral Bias in CRISPR Research

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    Blog Post
    ...Beroukhim and Boehm labs pulled data from more than 1000 CRISPR screens in DepMap to see if they could identify...Biomedical research has a long and well-documented history of racial bias, which still impacts and influences...
  12. Overcoming the AAV Size Limitation for CRISPR Delivery

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    Blog Post
    ...includes SpCas9, and shortern orthologs approximately 1000 amino acids in size. From the pool of shorter orthologs... 16, 2020 by Beth Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and ...packaging capacity of AAV (~4.7 kb). While this approach has been proven feasible, it leaves little room for additional...the number of targetable loci, and truncated Cas9 has much lower efficiency than its wild-type counterpart...amino acids, can also be packaged into AAV. NmeCas9 has the added advantage that it can be turned off by ...daunting, it’s actually a challenge that the field has overcome with split AAVs. In general split AAVs break...
  13. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...of template should have homology arms between 500-1000 bps. HDR efficiency with circular plasmid templates...applicable in DSB-induced genome engineering, it has biological importance for the repair of breaks that...directed repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made...
  14. Custom CRISPR Screens & the Green Listed Software

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    Blog Post
    ...design 5 guide RNAs (gRNAs) targeting each of the 1000 genes you’d like to investigate in your next CRISPR... population level, but each individual cell only has a maximum of one gene knocked out. In this genetically...containing up to around 200,000 different gRNAs. It has been suggested that researchers should use ~400X ...than gRNAs in a screen, to ensure that each gRNA has a good chance to contribute to the screen. In the... If you are lucky and find that a druggable gene has a central role in your system, you could majorly ...guest blogger Fredrik Wermeling! Fredrik Wermeling has a research group at the Centrum for Molecular Medicine...Medicine, Solna, Karolinska Institutet, in Sweden. He has a particular interest in using molecular tools to...
  15. Antibody Guide

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    Collection
    ...sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process may... immune response to it. Once the immune response has developed, the antibodies are collected, tested against...epitopes on constant regions of other antibodies. This has useful research applications, and is covered in the...exist as monomers. Each antibody class therefore has a valency, or number of binding sites available per...of antibodies multiplied by two, as each antibody has two binding sites. Isotype Description Configuration... must be able to detect if and where an antibody has bound. Signaling molecules , proteins that emit a...to avoid confusing data. Check that your machine has appropriate lasers and filters for each fluorophore...
  16. CRISPR 101: Multiplex Expression of gRNAs

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    Blog Post
    ... E. coli.” ACS Synthetic Biology 4(9) (2015):987-1000. PubMed PMID: 25822415. Find plasmids from this...multiple gRNAs on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll use mammalian...
  17. Fluorescent Protein Guide: Empty Backbones

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    Collection
    ...exctinction coefficient and quantum yield divided by 1000. pKa is the pH at which fluorescence intensity is...Addgene has assembled a collection of empty plasmid backbones with different fluorescent tags for you... that result in a diverse set of colors. Addgene has assembled a collection of empty plasmid backbones...
  18. Zinc Finger Consortium: OPEN Reagents

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    Collection
    ... for Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs that function in zebrafish... The Oligomerized Pool ENgineering (OPEN) method has been used to engineer ZFNs that function in zebrafish... Zinc Finger Consortium: OPEN Reagents (Kit # 1000000013 ) Depositing Labs: Keith Joung This kit consists...publication was a gift from Keith Joung (Addgene kit # 1000000013)" For your Reference section: Oligomerized pool...
  19. TALEN Plasmids and Kits

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    Collection
    ...respectfully). This next generation architecture has been shown to increase mutation induction when using...after the repeat domain, and C-terminal SV40 NLS and HA tags. Plasmid 47389 also contains the VP64 domain...double-selection of transfected cells, the system has successfully been employed for deletion mutagenesis...Sangamo)-FokI-ELD-Destination Golden Gate TALEN 2.0 1000000030 Yamamoto Lab TALEN Construction and Evaluation...
Showing: 1 - 20 of 65 results