...PubMed Central PMCID: PMC146043. 9. Nilsson, Stefan V., and Göran Magnusson. "Sealing of gaps in duplex DNA...gaps. For example, DNA assembly methods, such as Gibson Assembly® and NEBuilder® HiFi DNA Assembly, require...for a ligase? DNA ligases generally prefer fully Watson-Crick base-paired dsDNA substrates to those containing... base-pair size nearly indistinguishable from a Watson-Crick base pair, are joined with nearly the same...phosphate to the ligation). The structural/mechanistic reason for this differential is not known for certain,...allowed screening of all possible base combinations (Watson-Crick and mismatched) around the ligation junction....6 (2004): 757-764. PubMed PMID: 15582400. 2. Tomkinson, Alan E., et al. "DNA ligases: structure, reaction...

...I first became a manager. Know your people Each person is unique and should be managed accordingly.  I...role is not to do all the work yourself anymore. Personally I found this to be particularly challenging. ...important” tasks my schedule. This was bad for 2 reasons. First, it soon became obvious that I didn’t have...accordingly -  but think of it as an investment: once the person is trained you will have more time to focus on ...identify what to delegate and to whom (make sure that person has time for it!). Then schedule a meeting to discuss...Unless you’re delegating a recurring task to a seasoned report, there should be questions! Remember that...learned to embrace them after one ended with the person in front of me saying “I’ve been doing some of ...

...the Deverman lab at the Broad Institute and the Wilson lab at the University of Pennsylvania independently...unlikely receptor for PHP.B, and how the Deverman and Wilson labs determined LY6A was a receptor used by PHP.B... PHP.B CNS permissiveness Both the Deverman and Wilson labs determined LY6A, also known as stem cell antigen... role in viral susceptibility. The Deverman and Wilson Labs independently identified LY6A as a receptor...was a likely player in PHP.B permissiveness. The Wilson Lab also looked for the gene variant responsible...require galactose and AAVR to transduce cells. The Wilson Lab generated results similar to the Deverman Lab...’s transduction of 293T cells. In addition, the Wilson Lab also tested the ability of PHP.B to transduce...

... of your lab that you see in person, and helps improve your interpersonal relationships with people you...emailing documents to one person at a time. Just be sure there is a point person responsible for incorporating...replicates how you’d interact with team members in person, and improves overall communication.  Maintain...out if any milestones are missed. Spend time in person Presenting the results from my lab exchange...having collaborators spend time with each other in person. One possibility is to do a lab exchange, where...

...to our guest blogger Lydia Morrison from New England Biolabs. Lydia Morrison is a biochemist by training...This post was contributed by guest blogger Lydia Morrison from New England Biolabs. What is DNA assembly...multiple methods of DNA assembly available, including: Gibson Assembly®, BioBrick® Assembly, Golden Gate Assembly...Assembly, and NEBuilder® HiFi DNA Assembly. Gibson Assembly allows the production of scarless DNA constructs...opens the door for new opportunities in the field.  Gibson Assembly® is a registered trademark of Synthetic... Addgene Blog Learn More about Gateway Cloning Gibson Assembly and Other Long-Homology Based Cloning ...

...This post was contributed by guest blogger Chris Richardson, a Postdoctoral Researcher in Jacob Corn’s lab...and yet unanswered) question. Our recent paper (Richardson et al 2016) revealed some of the biophysical ...binding to the cleaved DNA duplex. Image from Richardson et al., 2016. We next wondered if we could...interaction with the Cas9-DNA duplex. Image from Richardson et al., 2016.   dCas9 enables sequence replacement...Results from our lab and others (Lin et al 2014, Richardson et al 2016, and Kim et al 2014) have shown that...transcribed sgRNA, and optional donor DNA. Chris Richardson is a Postdoctoral Researcher in Jacob Corn’s ...worked on this publication:         References 1. Richardson, Christopher D., et al. "Enhancing homology-directed...

...called GMAP (Gibson assembly-based modular assembly platform). GMAP is based on the Gibson Assembly method...Parton DL, Hanson SM, Rodríguez-Laureano L, Albanese SK, Gradia S, Jeans C, Seeliger MA, Levinson NM, Chodera...as individual plasmids shipped as agar stabs. Iverson, et al. ACS Synth Bio. 2015. PubMed PMID: 26479688...30-40bp overlaps). To expand the capabilities of Gibson assembly and make a more modular platform, the ...context-dependent human fitness genes, the laboratory of Jason Moffat at the University of Toronto has created ...Vectors Available at Addgene: Lab Plasmids Dan Carlson RCIscript-GoldyTALEN, pC-GoldyTALEN Tom Ellis...pTAL7a, pTAL7b Michal Zurovec pBlue-TAL Nathan Lawson pJDS Series   Superglue proteins with SpyTag...

...is. If the person makes time for you, as needed, this is a strong indicator that this person cares about...limited job opportunities in academia - the main reason most people enter postdoctoral training. Similarly... the postdoctoral mentor, and at least one more person that knows you and your research well. Who could...and skilled speaker. There's a more fundamental reason to give presentations - networking! People get ...unpublished information and reagents. Part of the reason I got my first faculty position was because the...institute about me, even though I had not asked this person to write a letter of reference. The group leader...not take time off when you are a PF. Here is my personal list:   Exercise - nothing dissipates stress ...

...Johnny Kung, Director of New Initiatives for the Personal Genetics Education Project (pgEd). Advances in...exacerbating existing inequalities and injustices. The Personal Genetics Education Project (pgEd), where I currently...conversations about both the benefits as well as the personal, social and ethical implications of genetics. ...cultural or religious affiliation, and ethnic or personal identity. We do this through a variety of activities...activities, including creating educational materials and lesson plans that are made freely available online; running...translating all these into social media and blog posts, lesson plans, or information briefs for policymakers. ...Kung is the Director of New Initiatives for the Personal Genetics Education Project (pgEd). He is broadly...

...labelled with horseradish peroxidase (HRP; Kristensson and Olsson, 1971), plant lectins (wheat germ agglutinin...24336151. PubMed Central PMCID: PMC4004172. Kristensson, K., and Olsson, Y. (1971). Retrograde axonal transport...difficult to accurately map network connections (Lo & Anderson, 2011). In the past two decades, several virus...PubMed Central PMCID: PMC4324159. Castle, M.J., Gershenson, Z.T., Giles, A.R., Holzbaur, E.L., and Wolfe...29, 363–365. PubMed PMID: 4107258. Lo, L., and Anderson, D.J. (2011). A Cre-dependent, anterograde transsynaptic..., J.S., Hyman, B.T., and McLean, P.J. (2009). Comparison of transduction efficiency of recombinant AAV... V., Thiry, I., Deroose, C. M., Mortelmans, L., Wilson, J.M., Debyser, Z., and Baekelandt, V. (2007). ...

...used to call copy numbers should be as small as reasonably possible and set using the variance seen within...negative for each reaction, math happens (this time, Poisson statistics), and the readout gives an absolute ... through concentration values, rather than by comparison to a standard curve, and most results can be ...7778-9_9. PMID: 29717442. Arya M, Shergill IS, Williamson M, Gommersall L, Arya N, Patel HR. Basic principles... S., Hollox, E. J., & Boon Peng, H. (2015). A comparison of assays for accurate copy number measurement...journal.pone.0116791 Providenti MA, O'Brien JM, Ewing RJ, Paterson ES, Smith ML. The copy-number of plasmids and ... Rachel Leeson...

...students who are unable to attend in-person interviews for a variety of reasons (logistical, financial, etc…)...visit in person, things like sickness or scheduling issues can and do interfere with those in-person plans...COVID-19 and standards for hybrid, virtual, and in-person events change seemingly daily (and are inconsistent...about the program and having the time to ask personalized questions might be difficult. However, an online...Even if you had planned a campus visit and the in-person portion got canceled, taking advantage of virtual...

...a price of a few dollars per reaction. By comparison, Gibson assembly can cost over $15.00 per reaction...In today’s post, we will focus on a high-level comparison of popular cloning methodologies, with a special...as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you first perform a...reaction if you purchase a pre-made mix. Nonetheless, Gibson assembly is a highly versatile and efficient way...propagating. The ligation-independent methods like Gibson and SLIC are also quite efficient; although efficiency...them are some of the homology based methods, like Gibson or SLIC, since one can pool the components and ...

... of a two-part interview with Vini Mani and Amy Gilson from Science in the News at Harvard University....your science, we sat down with Vini Mani and Amy Gilson from Science in the News, a graduate student organization...Podcast Many thanks to our guests, Vini Mani and Amy Gilson. Vini Mani is a graduate student at Harvard University...News, and member of the SITN podcast team.     Amy Gilson recently received her PhD from Harvard University...SITN podcast team. You can follow Amy on twitter @aigilson.   Special thanks to our sound engineer/producer...McFadden Learn about SITN's work at DayCon 2015 5 Reasons to Use Reddit for Science Communication ...

...changes in specificity than AAV-PHP.eB. For this reason, the Gradinaru lab anticipates that high-dose AAV-PHP.AX...happy transducing! This post was written by Rachel Leeson, with significant contributions from Cynthia Arokiaraj...Altermatt, M., Dobreva, T., Chen, S., Wang, A., Thomson, M., & Gradinaru, V. (2021). Deep Parallel Characterization...s41565-023-01419-x Deverman, B. E., Pravdo, P. L., Simpson, B. P., Kumar, S. R., Chan, K. Y., Banerjee, A....., Wang, Q., Katz, N., Buza, E. L., Bell, P., & Wilson, J. M. (2018). The Neurotropic Properties of AAV-PHP.B.... J., Bell, P., Raymond, A., Stanford, W. L., & Wilson, J. M. (2019). The GPI-Linked Protein LY6A Drives...journal.pbio.3002112 Jang, M. J., Coughlin, G. M., Jackson, C. R., Chen, X., Chuapoco, M. R., Vendemiatti,...

... is adapted from the 1979 Patterson Protocol. When published, the Patterson Protocol was shown to purify...DNA gel extraction method described in Kim and Morrison is nearly identical to that of commercially available...Purify DNA using glass filters. Image: Kim and Morrison, 2009. This method provides DNA yield and quality... kits for different purposes, with the Kim and Morrison method, you can just adjust the number of glass... protocol video library References Kim, Y., & Morrison, S.L. (2009). A Rapid and Economic In-House DNA...://doi.org/10.1038/s41598-018-30316-w  Zou, Y., Mason, M.G., Wang, Y., Wee, E.J., Turni, C., Blackall,...

...out their own novel experiments in semester two (Knutson et al., 2010a,b).      Fig. 1: Undergraduates...capabilities and laboratory and critical thinking skills (Knutson et al., 2010a,b). Classroom-based research projects...before a CURE trainee goes on to win a Nobel!   Jason Pellettieri is Professor and Chair of the Biology...Einstein Discovers America. New York./  Callahan, K., Peterson, C., et al. 2022. External collaboration results...planarians. Developmental Biology. 457:119-127. Knutson, K., Smith, J., Wallert, M., Provost, J. 2010a....and Molecular Biology Education. 38(5):317-323. Knutson, K., Smith, J., Nichols, P., Wallert, M., Provost...Education. 38(5):324-329. Kowalski, J., Hoops, G., Johnson, J. 2016. Implementation of a collaborative series...

...them for use in your lab. Structural Comparison Functional Comparison Advantages of Nanobodies How to Use...for a comparison of nanobody, HCab, and traditional IgG antibody structures. Functional comparison Strong...Nanobody Toolbox Find Nanobody Plasmids Structural comparison As the name suggests, nanobodies are smaller ...Large: Advantages of Antibodies One of the main reasons antibodies are great indirect immunodetection tools... antibody incubation. See figure 4A below for comparison of this one-step staining with nanobodies vs ...

...troubleshooting as advised by the technical support person. This delayed the progress of the project. After...familiarize yourself with technical support. Even having personal experience with technical support through the ...general positive attitude, and an open-minded personality. These traits will help you deal with any incoming...and there are also differences in customers’ personalities and their expectations. If you are willing to...is essential for success. For possibly this same reason, it can be one of the more constant and safe corners...simple way even if you don’t exactly know why. Personal and professional development in technical support...support gives you the opportunity to develop interpersonal and communication skills. Problem solving and...

...multiplex (Thellwell, 2000). See Table 1 for a comparison of features of several commonly used chemistries...sixfold variation in expression levels reported (Biassoni, 2014), so you’ll want to ensure that your reference...genes - and it is well worth your time to do one (Biassoni, 2014). At the bench As a highly sensitive technique... See Table 2, (adapted from Wong, 2018) for a comparison of different analysis methods.   Method ...defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. Table adapted... master. References and Resources  References Biassoni R; Raso A. (eds.), Quantitative Real-Time PCR:...doi.org/10.1016/j.bdq.2017.11.001.Bustin SA; Benes V; Garson JA; Hellemans J; Huggett J; Kubista M; Mueller ...