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...restraining further replication.   Figure 3: Comparison of replication regulation mechanisms of stringently...regulation! This post was originally written by Jason Niehaus in December 2015 and was updated by Emily...464.1998 Grimwade, J. E., Rozgaja, T. A., Gupta, R., Dyson, K., Rao, P., & Leonard, A. C. (2018). Origin recognition.../doi.org/10.1093/nar/gky457 Park, K., Han, E., Paulsson, J., & Chattoraj, D. K. (2001). Origin pairing...

...believe GMOs are safe for human consumption in comparison to 37% of the general public (5, 6). Similarly... the public (8–13). Facing the possibility that reason is losing ground in public discourse, many scientists...politicians to interpret it for them. Other popular reasons to march include: General inclusion of science...policy should have more weight than how any one person feels about it. And yet research shows that thoughts...

... make the best case for your candidacy. A major reason that a lot of candidates either get no interviews...understand your cover letter and research statement. My personal litmus test was whether my wife could explain ... please introduce yourself to me by email or in person. If you feel compelled to repay me in some way,...job searches. If you feel the urge to repay me personally, I will gratefully accept a very hoppy pint of...

...progeny are produced (Figure 1).  Figure 1: Comparison of the life cycle of a lentiviral vector with...integrase-deficient lentiviral vectors? There are several reasons why you might choose an IDLV over a lentiviral ...This blog post was written by Addgenie Rebecca Hutcheson.   References and resources References Banasik...PMC2835240 Wu Y. (2004). HIV-1 gene expression: lessons from provirus and non-integrated DNA. Retrovirology...

...fellow at the Morgridge Institute for Research in Madison, Wisconsin. It is commonly cited that approximately...enzyme function, protein interaction, etc. by comparison to non-phosphorylated or ‘wild type’ control ...dephosphorylation reaction can be tracked through the comparison of phospho:non-phospho species on PhosTag gels...fellow at the Morgridge Institute for Research in Madison, Wisconsin. She is particularly interested in the...

...nuclear- or DNA-binding proteins, you may need to sonicate your lysates in order to release the proteins ...you’re using chemical blockers, you can run a comparison of different formulations instead of concentrations...temperature, and/or buffer to increase lysis Add sonication step to protein lysis Splice variants Check...lysing buffer/adjust lysing buffer strength Sonication based lysing protocol Subcellular fractional...  Rachel Leeson...

...Addgene. Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, Crawford GE, Reddy TE, Gersbach CA...publication at Addgene. Maeder ML, Angstman JF, Richardson ME, Linder SJ, Cascio VM, Tsai SQ, Ho QH, Sander...Sander JD, Reyon D, Bernstein BE, Costello JF, Wilkinson MF, Joung JK (2013) Targeted DNA demethylation... this publication at Addgene. Mendenhall EM, Williamson KE, Reyon D, Zou JY, Ram O, Joung JK, Bernstein...

... appear within a range of wavelengths. For that reason, we speak of excitation and emission spectra. Both... happens due to a process called Fluorescence Resonance Energy Transfer (FRET).   Figure 1: Emission...fused to CD45, but is separately expressed. (The reason for having a separately expressed transduction ...difficult. I chose BV421 for the following two reasons: 1) The emission spectrum of BV421 is far enough...

...Single-chain variable fragment (ScFv) Figure 2: Comparison between the IgG antibody and scFv. Because...Variables (scFvs). Nanobodies Figure 3: Comparison of the Hcab and nanobody. Nanobodies are the...10.1016/j.ymeth.2017.01.009 Crivianu-Gaita V, Thompson M (2016) Aptamers, antibody scFv, and antibody...antibody Fab’ fragments: An overview and comparison of three of the most versatile biosensor biorecognition ...

... work, funding for science, and numerous other reasons. It is important to remember that we, scientists...Albany, NY, USA 42.6526°N 73.7562°W – Nicholas Thompson Over the last few years, there's been a disturbing...positive event that I hope happens again. Nicholas Thompson is a PhD Candidate studying Nuclear Science and...enforcement, and, although there was at least one person vocally protesting us, he was not threatening in...

...Many thanks to our guest blogger, Doug Richardson! Doug Richardson is the Director of the Harvard Center...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and... 2, 932-940 (2005). PubMed PMID: 16299478. 9. Richardson DS, Lichtman JW. Clarifying Tissue Clearing. ...

... something is working for you, then there is no reason to change it. But what if a kit isn’t working for...variety of manufacturers. It also includes a rough comparison of list price between these manufacturers, as...kits before deciding which one to buy. Table 3: Comparison of popular plasmid purification kit options ...blog post was written by Ashley Waldron, Hannah Dotson, and Nyla Naim. Resources Additional resources...

...for near-freezing temperatures, our first of the season snow flurries, dusting off the winter coats and... to #DeckTheLab with virus snowflakes from @CVRHutchinson #SciArt #STEAM #WinterWonderlab @Addgene Make...WFH space feel a little more festive this holiday season! It is that time of the year again: miniature ...

...PHP.eB Controls Wilson New viral vector pENN.AAV.EF1a.eGFP.WPRE.rBG AAV PHP.eB Controls Wilson New serotype... Rachel Leeson...

...Serotype Category Depositor  Notes pAAV_hSyn-SIO-stChrimsonR-EGFP-P2A-PdCO-miniWPRE AAV1, AAV5 Optogenetics...Biosensors Li New viral service pAAV-Ef1a-fDIO-ChrimsonR-tdTomato AAV9 Controls Jensen New serotype pAAV-CAG...resources!   This blog post was written by Rachel Leeson and Brian O'Neill.  More resources on the Addgene...

...vivo T cell Screens (FITS) approach (Milling and Markson et al., 2024), developed and deposited at Addgene.... Figure used with permission from Milling and Markson et al., 2024.   Proof of concept The FITS approach...cancer.   References and resources Milling, L. E., Markson, S. C., Tjokrosurjo, Q., Derosia, N. M., Streeter...

...description of the tasks I would be responsible for, in comparison to the other positions listed with a much less...flexibility was the ability to work either remotely or in person. This was useful to me as some days it was not ...their employees are feeling comfortable.     Lucie Wilson is a rising junior and bioengineering student at...

...note that these results could be biased for two reasons: 1) they are based on the US Occupational Employment...nonscientists, blur the line between professional and personal on Twitter. Or maybe scientist prefer to share...Tips: Online Social Networking for Scientists 5 Reasons to Use Reddit for Science Communication 10 Ways...

...Bioluminescence Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer...heterotrimeric G protein dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the...

...depolarization of R2 neurons using CsChrimson::tdTomato, an optogenetic tool (CsChrimson) attached to a fluorescent...Yang HH, Fisher YE, Wang IE, Ishida IG, Maimon G, Wilson RI, Clandinin TR (2019) SPARC: a method to genetically...

...mosaic virus Dr. Fiona Robertson, University of Zimbabwe, Zimbabwe Dr. Fiona Robertson (far left) is a Senior...potential for this drought tolerant plant, Dr. Robertson aims to make use of the CRISPR/Cas9 technology...

...scientific community, what other reasons do we like being a nonprofit? One big reason is that scientists and technology..., Eric, Gizela, Henry, Ina, Inna, Ishrat, Jane, Jason, Jessica, Joel, Jonathan, Justine, Karen, Kate, ...

...primers for In-Fusion Cloning Design primers for Gibson Assembly and NEB HiFi DNA Assembly  OligoAnalyzer...cloning methods, such as restriction enzyme cloning, Gibson assembly, or In-Fusion cloning.  NEB Ligation ...resources were added in September 2023 by Rachel Leeson.  Additional resources on the Addgene blog: ...

...matters! There are important societal reasons, self-interested reasons, and, given that science is largely...largely funded by tax-payers, there are reasons to feel obligated too, as experts on the subject brought out...

... mind from that session were words from Eric Hendrickson, loosely paraphrased in the tweet below. The... DNA for Enhanced CRISPR Genome Editing Chris Richardson and members of Jacob Corn’s lab describe how ...Yinan, Brian Ruis, Taylor Takasugi, and Eric A. Hendrickson. Genome Res 27(7) (2017):1099-1111. PubMed PMID...

...AAV9  pAAV-CamKIIa-ChrimsonR-mScarlet-KV2.1 124603  AAV9  pAAV-EF1a-DIO-ChrimsonR-mRuby2-KV2.1-WPRE-...affect DREADD trafficking and localization. The Sternson lab recently deposited PSAM AAV, which encode ...

...visualization tools using CRISPR In 2015, Thoru Pederson’s lab at the University of Massachusetts Medical...CRISPR-Sirius Recently, Hanhui Ma and the Thoru Pederson lab developed CRISPR-Sirius (Ma et al., 2018)....order to label two loci simultaneously, Ma and Pederson created another sgRNA containing the PP7 hairpin...

...science trainee will spend 4-7 years working for one person and with a group who all strive for a common big-picture...rotation, which reveals their working habits, personality, and gives them the opportunity to see if the...extreme openness” of the lab (i.e projects like the Personal Genome Project and open source software, hardware...

...comparable experimental results. Using PiggyBac transposons, the Woltjen Lab compared different variants ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at ...tool for use as a donor in FRET (fluorescence resonance energy transfer) and FLIM (Fluorescence Lifetime...1kb fragments are produced (see figure above). 3 Reasons The PSU Ladders Are Awesome Tools: They are comprised...

...error rate for long sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed a method using dCas9...error rates overall ranged from 0.3% to 10%, in comparison to the 62-90% error rate seen with Cas9 editors...barcodes for live cell clonal multiplexing by: Rachel Leeson Multiplexed images in live cells remains a challenge...

... variations in DR sequences. Figure 2: comparison of Cas 9 vs. Cas 12a expression cassettes when...0600-6. The Penn State Protein Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system...mammalian genome regulation and editing by: Rachel Leeson The large size of CRISPR-Cas effectors and fusion...

...Bradbury ARM, Trinklein ND, Thie H, Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B, Aldred SF, ...JoVE. https://doi.org/10.3791/54832  Lipman NS, Jackson LR, Trudel LJ, Weis-Garcia F (2005) Monoclonal ...

... Cpf1 for multiplex genome editing.  For a few reasons, Cpf1 is a simplified system for editing multiple...guide page for a review of Cas9.  For a brief comparison of Cpf1 vs. Cas9, see the table below. Table...expression vector.  See the graph in figure 2 for a comparison of editing frequency resulting from transfection...

... proto-oncogenes and disrupt their expression (Robinson, 1982). To add insult to injury, retroviruses ...immunogenicity low! References and Resources References Robinson HL. Retroviruses and cancer. Rev Infect Dis. 1982...Gene and Cell-Based Therapies in the New Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI...

... (fluorescent and luminescent) worlds Forster Resonance Energy Transfer (FRET) is the transfer of energy...which then emits it as light. Bioluminescence Resonance Energy Transfer (BRET) works the same way, except...tips below! This post was originally written by Jason Niehaus in June 2014 and was updated by Emily P....

... the genome. Mendenhall and Richard Myers of HudsonAlpha (where Mendenhall is also an adjunct faculty ...the function of the protein. Myers: We’ve done comparisons of our new system versus standard ChIP-seq for... for transcription factors, but there are many reasons you might want a system like this. It’s actually...

... Renaud J-B, Sallmyr A, Ruis B, Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin M, Brunet E...P, Burkhalter MD, Oh S, Qaqish BF, Moore DT, Hendrickson EA, Ramsden DA (2014) The fidelity of the ligation...

...want to cover during the year. Suggest that each person bring 3 topics and write them on a board to see... and meet regularly no matter what, even if one person can’t make it. One of the biggest group mentoring...meetings. Experiment with the process. Have each person write a Development Plan. Set concrete goals with...

... on what makes them that way. Is there a senior person whose career path you are interested in following.... Finding potential mentors is one of the many reasons to always be growing your network. If you don’t...need for your current position Understand your interpersonal challenges if they know some of the people you...

...decades to generate knock-out mice, but for various reasons it is not nearly as efficient in human stem cells...genetic background and do an apples-to-apples comparison of all those mutations. If you make induced pluripotent...backgrounds. It becomes more of an apples-to-oranges comparison. Addgene: Can you compare the TALEN kit to CRISPR...