...Though Cas9 is not a static molecule in any binding state, RNA loading stabilizes the closed conformation...pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo state (PDB ID 4CMP) with domains...the PAM is so flexible in this state that it doesn’t appear in the crystal structure (Figure 2) at all. ...Pro tip! X-ray crystallography cannot capture flexible or disordered regions, so crystal structures like...in this region. The crystal structure in Figure 3 shows the guide RNA stabilizing the DNA-binding protein...pyogenes Cas9 in the apo state (PDB ID 4CMP, top) compared to the guide RNA-bound state (PDB ID 4ZT0, bottom...engineering, and studying its structure can help us understand the parameters of CRISPR experimental design....

...the resources. Instead of creating a start up budget, create a start up list.  In this post we’ll help you...most start-up P.I.’s. It doesn’t have to be complicated, or difficult to create this list. Start BEFORE... much do you need to start your lab”. You can respond by saying, “The exact start up costs may vary greatly...Consulting, Inc. “How much do you need [for your lab start up]?” A hiring chair or dean will often ask a faculty...leads to poor outcomes; not enough funds to get started, wasting funds on unnecessary materials, or even... you successfully negotiate a start up award by giving you step by step instructions on creating a budget... in regards to the budget. Creating a budget We start by using your research specific aims. This is actually...

...chromatin and want to start sharing your plasmids through Addgene, don’t hesitate to start the process! Addgene...illustrations from Leah Bury of Microscopic Art. I started thinking about this problem when I recently attended...megabase away, or looking at the histone methylation status at a double-strand break that needs repairing. ... Pederson Phil Sharp Julie Ahringer Alexander Stark Bas van Steensel Richard Young Yi Zhang Leonard...H3.1 H3.3 HIST1H1E HIST1H2BB HIST2H2BE RBX1 STAG2 Xist Your favorite gene! SWI/SNF Genes:       ...

... and increase your lab sustainability. For my other posts on lab sustainability, check out my Recycling...the quick things above, try joining or starting a sustainability project. This could be: Setting up a ...This is the last post in a blog series on sustainable practices in the lab, where I’ll list some easy...unimportant. With so many big contributors to the sustainability problem, how can one little change make a difference...reusable water bottle instead of buying yourself a big stack of plastic cups. By initiating little changes in...lab Here are a few easy and fast things you can start doing right away to reduce your consumption in the...whole-cell extract from 2009. Join a newsletter to stay up-to-date on all the latest news and encourage ...

... Commercial DNA Ladder Penn State Ladder: Homemade Plasmid Prep Penn State Ladder: Commercial Plasmid ...undergraduate students led by Dr. Song Tan at Penn State developed the plasmids, pPSU1 and pPSU2. When restriction...$200-$500 for 1000 lanes worth of ladder. The Penn State ladders are significantly cheaper. When using a ...Your Agarose Gel Other Resources Read about Gel Stains on the BitesizeBio Blog ...

...to a few limiting factors.  To start, the average health science startup requires a great deal more capital... CEO of miLab, a Boston-based startup focused on empowering early-stage scientists. You can think of miLab...while becoming a health science entrepreneur at 22. Start with your network You've spent your life making ...the feedback we received from our network.  We started with the idea of a biotechnology investment fund...for such a fund is an impossibility at our early stage.  After each pitch we made, we considered the feedback...backgrounds (an essential component to any successful startup management team). Sreyan is a research associate... The world is against you.  As I mentioned, you start at a disadvantage in the health science space if...

...good ideas to a SynBio startup At some point, our ideas got pretty good. We started doing extracurricular...co-founders of Revolution Bioengineering.  Last year we started a company. Revolution Bioengineering is two responsible...working in a new technical discipline that nobody understands (synthetic biology), building a product no one...a plant that could sense and respond to novel substances. We both had an applied science bent, so the ...postdoc in order to rebuild my resume, so I had started to entertain other options. Keira had it a little...occasionally thinking about how we could bypass the establishment hiring system altogether. We had a lot of ideas...and dissected with equal enthusiasm. It may have started as idle conversation to pass the time in a job ...

...interview with Crystal Shih Podcast breakdown Intro Chapter 1: Who is Crystal Shih? Crystal's educational...Gray at Caltech, Crystal moved on to a Postdoc in the chemistry department at MIT. Crystal now works on protein...skills. Outro Many thanks to our guest, Crystal Shih. Crystal Shih leads project and scientific teams ...Crystal Shih is an Investigator III at Novartis, a multinational pharmaceutical company based in Switzerland...therapeutics at Novartis - listen to learn more about Crystal’s exciting career path, to gain insight into industry...to succeed in your own career. One pro tip from Crystal - be agile and willing to move on to new projects...how delegating can empower people Chapter 3: How Crystal advanced at Novartis, and how to prepare yourself...

...three larval stages where larvae molt in order to continue growing in the next larval stage. After the ...remain stable as homozygous alleles, others cannot. Geneticists need a method for maintaining stable lines...three-part series. We’ll first discuss how to get started on fly work in this post. The second post will ...the bees when it comes to fruit flies Drosophila start life as fertilized eggs that develop into embryos...the third larval stage, Drosophila will pupate, which takes about one week from the initial deposition of... visible dark spot. Pupae begin eclosion at the start of the day and throughout daytime. They mostly stop...through their translucent bodies. This is the ideal stage to select for females because you can be certain...

...Recombinase-based state machines (RSMs), which take on a particular DNA-sequence configuration (state) based on...effects on the state cannot be undone and also limit the set of possible downstream states that can be achieved...order of inputs results in only one final state, then the state of the RSM will tell you precisely what ...might one read the state of an RSM? The most straightforward way to read the state of an RSM is to sequence...is to interrogate state with qPCR. The re-arrangement of DNA segments in each state of a register enables...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...green fluorescent proteins in each state. The three methods of state interrogation mentioned above (Sanger...

...have been developed, which differ in the number, stability, and brightness of the fluorescent proteins used... (Cai et al., 2013), incorporates the highly photostable fluorophores mOrange2, EGFP, and mKate2 (red)...truncated, non-fluorescent form of YFP as the default state of the transgene such that recombination is necessary...not undergo recombination can be detected by immunostaining of the truncated YFP. In Thy1-Brainbow mice...2011) labels Drosophila neurons at the neuroblast stage, meaning that neurons of the same lineage will express...injecting a CMV-Brainbow construct at the embryonic stage. This technique could be used to target Brainbow...labeled by injecting Brainbow at the single-cell stage. Zebrabow, or “zebrafish brainbow,” is a collection...

...In 2008 the Quake Lab at Stanford University became interested in exploring biological dark matter – ...Preparations ahead of time included contacting YNP staff to arrange permits to visit places not accessible...voltmeter on top of the battery box to monitor charge status and a disconnect switch between the battery bank...September we arrived in YNP just as the weather was starting to turn chilly at the 8,000 foot elevation. Due...environmental samples Since the high temperatures and unstable ground around these hot springs make them extremely...telescoping poles to collect water samples from a distance. At each collection site, we took GPS coordinates... collected small water and sediment samples, stabilizing them in ethanol and/or glycerol, since there ...

...example, I am not that interested in meeting movie stars, but when I met the astronaut Jim Lovell I was speechless...work one floor above mine in the lab of Dr. Jack Szostak, who went on to win the Nobel Prize. Dr. Doudna...whiteboard. The final image is pictured here…she started from a blank white slate and developed this over...paper in Science.  Interestingly, this project started with their labs studying the mechanisms of RNAi...

...entrepreneur must be OK with the constant fear of failure at all stages in the startup". To gain insight into "...team was unable to secure enough money to sustain a startup, he went to work for Flagship Ventures – a... his energy and effort into making the startup self-sustainable. Within a year from this decision, Mike...Development & Operations. Starting a biotech company When and how does a startup become "successful"? First... budding startup, I asked Mike to outline 3 pieces that are essential to the biotech startup puzzle: ...Assembly Standards Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)    ...occasionally pondered translating my research into a startup company. However, like many like-minded people,...

...into two stages: Dedifferentiation - Conversion of adult somatic cells into the pluripotent state. Redifferentiation...differentiated somatic cell reverts back to a pluripotent state, which then proliferates and is redifferentiated...lineage, without dedifferentiating into a pluripotent stage. Cellular reprogramming The reversal of a differentiated...differentiated cell type to an undifferentiated state and then redifferentiation into the cell type of choice...differentiated cells of choice. The dedifferentiation stage involves overexpression of four reprogramming factors...differentiated somatic cell to revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate...reprogrammed to iPSCs successfully [10-12]. The next stage of reprogramming consists of redifferentiation of...

...replicate while maintaining the undifferentiated “stem” state and, Their potency or capability to differentiate...cells found in the zygote through to the morula stage are totipotent [2-4]. Pluripotency: When a cell ...

...informative introduction for those who are just starting their careers in the lab and even those who might...

... panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (pink) to indicate...Image from Isaacman-Beck et al., 2019. Get started with sparse labeling Isaacman-Beck et al., bioRxiv...bioRxiv 2019. https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast...transcriptional reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X CDRE repeat. In addition... that this method is compatible with dead cell staining using propidium iodine (PI), allowing for calcineurin...Truncated TALE-fluorescent protein fusions as a DNA staining dye Article contributed by Jennifer Tsang ...generated fluorescent protein fusions with tTALE to “stain” DNA. TALE proteins can bind specific DNA sequences...

...their promising research projects.   Understanding drug resistance mechanisms of Mycobacterium tuberculosis...interested in understanding the mechanisms behind Mycobacterium tuberculosis resistance to pyrazinamide...targets. An early and accurate detection of PZA-resistance is necessary to improve tuberculosis care and...Sheen aims to identify new genes related to PZA-resistance by using different approaches, including CRISPR...Her research should provide us with a better understanding of PZA action and could potentially stimulate...diagnostic methods and improved therapies for PZA-resistant Mycobacteria tuberculosis. Investigation of multispecies...this condition are urgently needed. Therefore, understanding neuronal networks, their interactions and information...

...chemistry behind Gram’s crystal violet stain and the ion interactions that cause the stain retention in gram...positive bacteria trap the crystal violet stain within their membrane, while the stain is washed from the gram...realized that certain stains were retained only in some species of bacteria. The stain colored one type, ...Gram’s discovery, adding a second stain, safranin, as a counterstain to help visualize the bleached bacteria...Procedure The stain is a pretty simple process. First, flood the heat-fixed smear with the crystal violet stain... bacteria will be stained blue or purple, and gram negative bacteria will be stained red or pink, as shown...mixing the stains and solutions used, check out these resources.   Bruckner, M. Gram Staining. https://...