...commonly used in optogenetics: 1) electrophysiology and 2) behavioral experiments. I’m not an electrophysiologist...choice in the brain region you wish to stimulate 2) Fabrication and implantation of the optical fiber...thoroughly elsewhere (here and here for instance) [2], [3]. Karl Deisseroth of Stanford University , one...reviews for a discussion of the different variants [1, 2, 4]. And, of course, the best way to learn about the...monitor expression at various time points (e.g. every 1-2 weeks over 8 weeks) in order to determine the optimal...(4):251-66. doi: 10.1038/nrn3171. PMID: 22430017. 2. Fenno L, et al. The development and application of...analysis of microbial opsins. Nature methods. 2012;9(2):159-72. doi: 10.1038/nmeth.1808. PMID: 22179551; ...

..., AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors contain type 2 ITRs, which comes from...differentiated, non-dividing cells.   Figure 2: DNA rescue and transduction of a conventional single-stranded...quantitative PCR-based method for the titration of AAV-2 vector stocks."Molecular Therapy 6.2 (2002): 272. ...Guerin and Susanna Stroik. Didem Goz Ayturk is a Postdoctoral Fellow in Connie Cepko’s Lab investigating neuronal...

...blood are reprogrammed immediately after collection [2]. PBMCs display unique epigenetic signatures that ...PMID: 27882265. PubMed Central PMCID: PMC5118044. 2. Kim, Y., et al., The Generation of Human Induced ...pCXLE-hUL 27080 L-MYC and LIN28 pCXLE-hOCT3/4-shp53-F 27077 OCT3/4 and shRNA against p53 pCXWB-EBNA1... is the expression of the reprogramming factors- OCT3/4, SOX2, KLF4, and C-MYC [1]. For the derivation...mixture of two vectors that encoded six factors - OCT3/4, SOX2, KLF4, C-MYC, LIN28 and SV40 large T antigen...Mukherjee. Kusumika (Kushi) Mukherjee is currently a postdoctoral fellow at Massachusetts General Hospital with...

...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...contributed by guest blogger Jessica Polka, a Postdoctoral Research Fellow with Pamela Silver.  Most types...with their environment.   Jessica Polka is a Postdoctoral Research Fellow with Pamela Silver in the department...

...mammal called alpaca could neutralize the SARS-CoV-2 virus. But while the early results were promising,...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a specific...Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert’s suitability and availability...IgnacioAmigoH     María Hernández-Sánchez is a postdoctoral researcher at Instituto de Investigación Biomédica...

...therapy had fallen - plummeted really -  from grace 2 years prior with the death of Jesse Gelsinger in an... the whole field to a grinding stop and only now, 2 decades later, is it finally back in the limelight...the other hand, have broad tropism in the eye. AAV-2, the first identified serotype and the one used in...reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to be the next...amaurosis. This is the first of what I, scientists, doctors and patients hope are many more clinical successes...

...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...Primer design Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification... for each type of modification is shown in Figure 2, and is discussed in more detail below. The main requirement...Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2. Bubeck, Peter, Monica Winkler, and Wilfried Bautsch...Laboratory of Molecular Biology. Jake Watson is a postdoctoral researcher in neurophysiology at the MRC Laboratory...

...FLP and Cre for additional control   Figure 2: Expression of a morphological marker (eYFP) and synaptic...Sternson. 2008. “A FLEX Switch Targets Channelrhodopsin-2 to Multiple Cell Types for Imaging and Long-Range ...PMID: 18614669. PubMed Central PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart....The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to DNA via a Phosphotyrosyl...guest blogger, Katrin Michel! Katrin Michel is a Postdoctoral Researcher at MIT and is fascinated by the question...

... will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models reinforcement...spend most of its times in the opposite chamber) [2]. Laser activation is usually controlled automatically...two different ways 1) your hypothesis is wrong or 2) there is some problem with the technical implementation...PMID: 21716290. Pubmed Central PMCID: PMC3775282. 2. McDevitt, Ross A., et al. "Serotonergic versus nonserotonergic...University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne Kreek at the...

...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression...Patrick Miller-Rhodes, a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center...Miller-Rhodes is currently a Ruth L. Kirschstein NRSA Predoctoral Fellow at University of Rochester Medical Center...

...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https...the insectspodoptera frugiperda (lepidoptera; noctuidae). In Vitro, 13(4), 213–217. https://doi.org/10.1007...

...Public Release Event scheduled for March 4th from 2:30-4:00 pm ET.  You can learn more about the report's...professional development as core components of predoctoral and postdoctoral training. A grant from the National ...

...symposium on the future of the scientific endeavor October 2-3, 2014 Boston University Jacob Sleeper Auditorium...article - you should join Dr. Bourne in Boston on October 2nd when he will be the keynote speaker for the...

...native enzyme, which are summarized in Table 2 below.  Table 2: Properties of native luciferases Organism...None Yes Luminous Japanese ostracod(Cypridina noctiluca)Sea firefly(Cypridina hilgendorfii) 465 62 Cypridina...

...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...then moved to France where she worked as a post-doctoral fellow on developmental biology at the École Normale...

...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...candidates for further development. They inserted an octet of MS2 hairpins into the gRNA tetraloop and went...

...Interview with Stem Cell Researcher Darrell Kotton 2. Stem cell scientists care about patients As any grad... Yamanaka explained that it took 50 people nearly 2 years to create the first stem cell line that could...differentiated cells simply by expressing four proteins Oct4, Sox2, Klf4, and cMyc (the so called OSKM factors...

...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...notebooks you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn...Management Software Resources on Addgene.org Find Protoctols for Your Research Catch Up On Your Molecular ...

... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your...sponsorship) then he or she can start working in October of the same year. The Industry H-1B visa is not...research institutions are willing to sponsor postdoctoral researchers for the EB-1B using the concept ...

...represent a favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive...cell 30.1 (2016): 147-160. PubMed PMID: 27411589. 2. Todoric, Jelena, Laura Antonucci, and Michael Karin...Relationship” by our guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University! Cancer therapeutics...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor...Nucleic Acids Research, Volume 28, Issue 19, 1 October 2000, Pages 3752–3761, https://doi.org/10.1093/...

...and scholarships in different subject areas!    2. Resume and cover letter know-how  Tailor your resume...4. Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch...Pierre! Roodolph (Roo) P. St Pierre is currently a doctoral candidate in the Chemical Biology Program at Harvard...

...is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation...working group (National Academy of Sciences. “The Postdoctoral Experience Revisited” 2014 Appendix B p93-95...

...efficient knockout (KO) of genes in immune cells; (2) is not immunogenic, a necessity for in vivo studies... Find FITS plasmids here! Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard...

...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve... tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine)– Nonnatural amino acids can be ...

...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...www.youtube.com/watch?v=sx1uDYSfINA. When you go to the doctor, it will very likely take time to determine if ...

...interferon gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of...contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University Cancer is one...Subhadra Jayaraman! Subhadra Jayaraman is currently a doctoral candidate at Binghamton University. She works ...

... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...resources with examples, like BioRad’s Western Blot Doctor page, or robust discussions, like ResearchGate’...

...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...Circadian Rhythm and Melatonin for the Timing of Nocturnal Courtship Vocalization. The video summarizes the...

...information gathered by Postdoctoral Associations around Boston - From Panel Session 2: Defining the Postdoc...research community to develop a plan to make postdoctoral pay/benefits commensurate with work training...a postdoc as “An individual who has received a doctoral degree (or equivalent) and is engaged in a temporary... staff scientists. This results in a glut of postdoctoral trainees who, even though they are aware that...become actionable. Additional Resources The Postdoctoral Experience Revisited. PubMed PMID: 25590106....

... of my favorite molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique...deposited plasmids on this trip. Here’s to hoping that October is even more awesome than September! Tyler J. Ford...

...citizens enter into in order to protect their rights (2). Less formally, governments are the organizations...science education and literacy that this changes.  2) Lumping all scientists (and consequently their results...active research scientists (scientists who hold a doctorate degree, are currently working full-time, and have...

...), 1668. https://doi.org/10.1038/s41467-019-09656-2 More resources on the Addgene blog CRISPR101: Validating... X-CHIME plasmids here!   Marty LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard...

...double strand break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...post was originally written by Jennifer Tsang in October 2019 and updated by Emily P. Bentley in December...

... stabilization of unknown bacteria. Figure 2: Christina Mork collecting samples at the hot spring...Thermus aquaticus bacterium – was discovered at Octopus Hot Spring at YNP, and since then, many researchers...

...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...And, yes, your partner CAN take the kids to the doctor without your help. Your partner is equally responsible...eight-year-old daughter vehemently reminded her doctor that she was allergic to penicillin – I am much...

...externally fertilized, transparent progeny (Figure 2). These features give it a leg up (or a fin up?) over... post was contributed by Katherine Rogers, a postdoctoral researcher at the Friedrich Miescher Lab of ...Max Planck Society. Katherine W. Rogers is a postdoctoral researcher in Patrick Müller’s group at the ...

... and whether I would be able to carry out a yeast-2-hybrid library screen; instead I thought they would.... I had just started month 8 of my 3 year Post-Doctoral contract at the University of Cambridge and knew...

...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...Bioinformatics from Boston University, and did postdoctoral work at Harvard Medical School and the University...

...two visible blue or magenta chromoproteins (Figure 2) (Liljeruhm et al., 2018). This system is a nice confirmation...This post was contributed by Jacob Lazarus, a postdoctoral researcher at Harvard. There’s an astounding...is now an attending physician. He performs post-doctoral research in the lab of Matthew Waldor at Harvard...