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...removed to leave you with DNA. This DNA is then washed, and either eluted from a fixed surface or precipitated...glass particles are pelleted by centrifugation and washed before DNA is eluted with water or TE. The Bishop...adapted from the 1979 Patterson Protocol. When published, the Patterson Protocol was shown to purify DNA...

... incubator). This means any other scientist who wishes to use these cells can see their passage number...immortal life of Henrietta Lacks. New York: Crown Publishers, 2010. 2. Gold, Michael. "A conspiracy of cells...Additional Resources ATCC Animal Cell Culture Guide Thermofisher Guidelines for Maintaining Cultured Cells Invitrogen...

...(Gootenberg et al., 2017). This system, dubbed SHERLOCK (depicted in figure 1) has been used to differentiate...applications of CRISPR. Since this post was originally published, we've also seen the new development of Cas13b...al., ‘Point-of-care testing for COVID-19 using SHERLOCK diagnostics.’ medRxiv 2020 May 8. Pubmed PMID:...

... Originally published Jul 14, 2015 and last updated Sep 16, 2020 by Beth Kenkel.  CRISPR genome editing...Cas12b and CasX. While there are no papers yet published that deliver these Cas proteins with AAV, at just...0908-x  Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O (2014...

...maintaining accuracy.  This post was written by Alyssa Shepard, with significant contributions from Alyssa Cecchetelli...5467. https://doi.org/10.1073/pnas.74.12.5463  Shendure, J., & Ji, H. (2008). Next-generation DNA sequencing... Alyssa Shepard...

... is a loop in between secondary structure (beta-sheets or alpha helices). An additional requirement is...be generated with a transposon-based strategy (Sheridan, 2002). Avoiding protein over-expression artifacts...PMID: 18454154. PubMed Central PMCID: PMC2853173. Sheridan, Douglas L., et al. "A new way to rapidly create...

...depending on the structure of the TO derivative (Dolgosheina et al., 2014). While 10 derivatives of TO exist...ingredients in the aptamer soup In addition to these established RNA aptamer fluorophores, more are in development...29440634. PubMed Central  PMCID: PMC5811451. Dolgosheina, Elena V., et al. "RNA mango aptamer-fluorophore...

...repressed more   LacZ β-galactosidase activity abolished   lacY Lactose permease inactivated, lactose...plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability, reduced recombination...r-m+)  MG1655    Wild type K-12 strain; first published sequence of an E. coli K-12 strain. F- λ- ilvG...

...unlock” and infect specific cells. This protein shell packages the vector’s genetic material, and variations...the first paper identifying a binding receptor published this year (Dhungel et al., 2025)! It can transduce...contributing their expertise! This post was originally published in January 2025 and updated in August 2025. References...

...teams: Watson, Crick, & Franklin, Meselson-Stahl, Hershey-Chase...Who was I to dispute history? And boy did...organization on the planet, in which case whoa! Mission accomplished! One example of a mission-based team in academia...myth. The original Dr. Frankenstein from Mary Shelley’s novel worked alone, but even the movie makers ...

... buffer Microcentrifuge tubes BCA Assay, Thermo Fisher 23227 β-mercaptoethanol 4X protein loading buffer... ladder Ethanol iBlot 2 PVDF Mini Stack, Thermo Fisher IB24002 20X TBS Tween-20 Nonfat milk powder 96-...deionized water 3x . Shake the gel gently between washes to remove residual water. Load the gel into the...

...Lumio Green and Lumio Red from Invitrogen (now ThermoFisher Scientific). These reagents are nonfluorescent...is that binding is covalent, so free dye can be washed away. In addition, labeling contrast can be improved... developed, so don’t be afraid to try novel, unpublished combinations of labeling strategies. FRET experimental...

...around the same time. (The papers were actually published in the same journal on the same day.) This team...lot of terminology in this post! Here’s a quick refresher if you ever lose track of which acronym is which...10.1038/s41556-023-01342-2 Koblan, L. W., Arbab, M., Shen, M. W., Hussmann, J. A., Anzalone, A. V., Doman,...

...CRISPR editing world. The approach, originally published in Nature, is a “search-and-replace” genome editing...Yang, W., Leith, A., Daza, R. M., Martin, B., & Shendure, J. (2022). Precise genomic deletions using paired...s41587-021-00868-w Nelson, J. W., Randolph, P. B., Shen, S. P., Everette, K. A., Chen, P. J., Anzalone, ...

...optimized primers that may work better than the published ones, so if/when you use HercII or Kapa Hifi the...all FAQs from Le Cong on June 24, 2025 by Alyssa Shepard.   References and Resources References Beumer...Additional resources on the Addgene blog CRISPR Cheat Sheet Build Your CRISPR Vocabulary CRISPR 101: Off-Target...

...transcription start site. In STARR-seq, randomly sheared genomic libraries are cloned downstream of a minimal...simple and complex DNA constructs Have you ever wished you could design a gene knockdown or overexpression...characterize positive hits. This work has been published in Cell. The Moffat laboratory maintains a website...Center has kindly constructed and deposited an unpublished set of Zika gene plasmids to Addgene’s repository...

...to Cas9. All of this is accompanied by new or refreshed figures made using BioRender.com. Figure... Alyssa Shepard...

...you ever thought - Is there an easy way to find published plasmids? Do you spend a lot of time in PubMed...to set-up emails to alert you when papers are published on your favorite topic. If you have any other...

... serotype pAAV-hDlx-Flex-dTomato-Fishell_7 AAV2 Controls Fishell New serotype pAAV-mDlx-NLS-mRuby2...

...A newly established all-in-one vector construction system for CRISPR/Cas9-mediated multiplex genome engineering.... “However, there had been no systematically established way of making an all-in-one vector for multiplex...