...Molecular Cloning Techniques References Bell AD, Usher CL, McCarroll SA. Analyzing Copy Number Variation...10.1007/978-1-4939-7778-9_9. PMID: 29717442. Arya M, Shergill IS, Williamson M, Gommersall L, Arya N, Patel ... Curr Protoc Hum Genet. 2014;80:7.21.1-7.21.. Published 2014 Jan 21. doi:10.1002/0471142905.hg0721s80 ...

...Originally published Dec 7, 2017 and updated Jul 2, 2020. Promoters may be the star of gene regulation...Elements) method. The original CAPTURE method was published in 2017 and addressed many of the drawbacks of...where they are physically located. DNA is then sheared into pieces small enough to be pulled down via ...

...techniques. More and more CRISPR tools are being published each month, making CRISPR a great choice for your...to consult the literature to see if anyone has published a protocol that would work for your system. Addgene... are a few plasmids at Addgene that have been published as CRISPR testing tools: Traffic Light Reporter...

...can be eluted once the other materials have been washed away (Figure 1).      Figure 1: The steps...while the media is aspirated. The beads are then washed, followed by antibody elution. Whatever method ... strong ones, consider increasing the number of washes performed during your harvest. Validation Recombinant...

...its various derivatives, biological samples are enmeshed in a gel of a highly absorbent material that is...full IgG secondaries, can improve signal strength (Sherry and Stiles, 2022). Fourth, signal continuity can...20210110. https://doi.org/10.1098/rsta.2021.0110  Sherry DM, Stiles MA (2022) Improved fluorescent signal...

...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...

...High-throughput cloning, in a nutshell, is the systematic combination of different genetic sequences ...of coding sequences Sladitschek and Neveu, 2015; Shetty et al., 2008; Anderson et al., 2010 Among the...1961. March;236:864–875. Pubmed PMID: 13790780. 5. Shetty RP, Endy D, Knight TF. Engineering BioBrick vectors...

...This post was contributed by Max W. Shen from MIT, Alvin Hsu Harvard University, and David R. Liu from...uncertainty. Many thanks to our guest bloggers Max W. Shen from MIT, Alvin Hsu Harvard University, and David... the Broad Institute and Harvard University. Max Shen is a Ph.D. candidate at MIT. His research uses applied...

... 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...

...(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:...

... 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...

... 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...

...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 ...

...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, ...

...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...

... 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...