...extraction solution per well and resuspend. Continue to step 7.3 for suspension cells. For adherent cells, ...CRISPR Cloning Anneal and phosphorylate oligos. Resuspend oligos at a concentration of 100 μM in ddH 2 O...detail for murine erythroleukemia (MEL) cells, a suspension cell line. The culture medium in all steps consists...strategies for each cell type. While MEL cells are suspension cells, instructions for adherent cells have ...Ensure there are 2 x 10 6 cells per CRISPR pair. Resuspend 2 x 10 6 cells in 100 μl of electroporation solution...validation (see step 6 ). NOTE: This protocol is for suspension cells. Adherent cells can either grow as individual... gDNA from parental and bulk sorted cells by resuspending parental and bulk cell pellets in 50 μl of DNA...

...and non-coding RNAs in zebrafish - Tatjana Sauka-Spengler Lab Lineage Tracing Cell lineage tracing in multicellular...drivers in vivo. Biotagging toolkit - Tatjana Sauka-Spengler Lab. A genetic binary system for cell- and compartment-specific...

...millions of patients results in billions of dollars spent each year. Unfortunately, the causes of most neurodegenerative...

...the growth medium and can either be used while suspended in the original media or isolated and placed in...

...2A-mCherry-FKF NLS-Cre, mCherry T7 Bacterial Sauka-Spengler 119971 GAG-CRErec GAG-Cre fusion; for the production...

...the growth medium and can either be used while suspended in the original media or isolated and placed in...

...but you’ll only spend half of each day on it. The other half of each day you can spend accomplishing small...’d be too late to meet up, and I’d just end up spending more time in the lab. Obviously I didn’t have ...didn’t realize it until it was too late. I didn’t spend enough time thinking about the possibilities and... moderately useful tasks at the expense of time spent on the long and highly useful tasks. The quick accomplishments...complete the entire experiment, but you’ll have to spend the entirety of each day doing the experiment. The...had general goals for my experiments, but didn’t spend enough time thinking about when or how long it would...

...budget were proposed. I spent a lot of time asking why. As scientists we spend years being taught to communicate...the solutions they promote help motivate you to spend a little time advocating for science and getting... New Yorker, so it’s not easy to convince me to spend a Saturday in the tourist-trap nightmare that is...disconnected from the entire scientific process.  I've spent a lot of time debating how to fix this.  As scientists...W - Erin Golden Hi, my name is Erin and I have spent the last 15 years training as an academic scientist...

...arm (safe). Mice with higher IL-1 beta production spent more time in the open arm and were less risk averse... dark (protected) area. When measuring the time spent in a particular part of the apparatus, it turned...turned out that the IL-1 beta overexpressing mice spent more time in the risky bright place, driven by their...behavioral despair, and the more time the animal spends in the passive mode, the stronger is the depressive...swim test as seen by increasing the ratio of time spent actively swimming versus passive floating. In the...

... at Addgene as an Outreach Scientist. I used to spend my days (and often my nights :D) engineering E. ... Pamela Silver’s Lab (plasmid page here). Now I spend much less time wrangling bacteria and instead help...in my position as an Outreach Scientist where I spend a large amount of my time meeting with and talking...on a plane back to Boston from Kentucky where I spent a couple of days talking to researchers about their...

...A scientist-in-training will spend 10 or more years with a small number of formal advisors learning how...determining whether an advisor will be a good mentor: Spend time with the potential advisor and don’t be shy... in the lab, if possible. Attend a lab meeting. Spend some time hanging out with members of the lab. It...sign if the PI does not welcome you to visit and spend time with everyone. Additionally, if you are looking...

...can be a major decision. A science trainee will spend 4-7 years working for one person and with a group...gives a 1 hour talk about his or her interests, and spends one-on-one time with the current members to share...solid foundation on which to thrive during time spent as a graduate student or postdoc. Although the chosen...completely happy at work all of the time, the goal is to spend as much time in the "zone" as you can. You know ...

...protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids ...

...experimental limitations and considerations? I recently spent two months in the lab of Antonello Bonci, Director...The technique is non-trivial; you don’t want to spend months doing surgeries and making fiber optic cables... method of identifying bregma but it is best to spend the time practicing your injections until they are...accurate and reproducible. I also highly recommend spending the money on a high quality stereotaxic frame ...

...University, you are expected to spend at least one semester (the majority spend two to three) working in your...of my six month employment. In the beginning, I spent a lot of time shadowing members of my team to learn...

...Addgene, I had spent most of my junior year stressed out applying for summer programs. I spent one day at ... before high school senior year would have been spent within a lovable community that has allowed me to...

... and reaching out if any milestones are missed. Spend time in person Presenting the results from...circumstances permit, think about having collaborators spend time with each other in person. One possibility ...possibility is to do a lab exchange, where researchers spent time in each other’s labs. This is an opportunity for...

...to make a plasmid? Or how much time you have to spend cloning before you can start your experiment? What...the most valuable component in science – the time spent by the scientists designing and performing the experiments...but we can assume it’s at least equal to the $155 spent on reagents. Five ways to decrease plasmid production...

...this method, beads are incubated with the cell suspension. Antibodies bind ligands on the beads and a magnet...Grazzini E, Durocher Y. Transient Gene Expression in Suspension HEK293-EBNA1 Cells. Methods Mol Biol. 2018;1850...on magnetic beads from very high cell density suspension. Biotechnol Bioeng. 2021 Sep;118(9):3499-3510...

...the mix. Thus, our founders and early employees spent this remaining time by elaborately pranking each...play hard.” Part of the reason we sincerely like spending time together is that when it’s time for fun, ...more manageable when you can enjoy the people you spend 40+ hours with each week. In conclusion - get out...