...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project. Gymnotiformes and Mormyridae...evolution in immunology, neurobiology, and more. Prochlorococcus (Penny Chisholm) This cyanobacterium is the...need to grow to get a recombinant because this process is expensive. They are using a cool fluorescent...

...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit...and eukaryotes, which use a binary replication process whereby one cell copies itself and “splits” into...genome with proteins central to the replication process removed. This means it can deliver a genetic payload...

... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...doesn’t, and trying all over again. Here’s that process!   Figure 1: The Design-Build-Test-Learn...gear, and walking everyone through the recording process was going to be time consuming and difficult. Finding...in the future, but we were finally refining the process down to a place I could be happy with it. Cycle...

...delicate axonal and dendritic processes not previously visible with Brainbow-1 and -2. The general structure ...Brainbow mouse neural labeling system. Brainbow-1 and -2 have been further refined to create Brainbow 3.2. ...M-CFP expression; these deletions are defined using 2 loxP variants (only identical loxP sites can mediate... each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained a “default” state; for example, Brainbow...

...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...Getting your RQ value isn’t the end of your analysis process. You’ll still need to do statistical analysis to...Gene Expression Analysis During iPS Reprogramming Process. Sci Rep 8, 8716 (2018). https://doi.org/10.1038...

...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...vectors Learn about Addgene AAV Quality Control Process Resources on Addgene.org Find ready-to-use AAV...

... is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...biology98.3 (1975): 503-517. PubMed PMID: 1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new...recording DNA abundance throughout the amplification process, rather than just at the end as in standard PCR...became the workhorses of molecular biology." Proceedings of the National Academy of Sciences 102.17 (2005...restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy of Sciences 68.12 (1971...sequencing with chain-terminating inhibitors." Proceedings of the National Academy of Sciences 74.12 (1977...

...phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip..., accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/Cas moieties carry out these functions... also disrupted to elucidate their role in the process. The results of these studies led scientists to...sequence. Separate Cas proteins then express and process the CRISPR loci to generate RNA, which guide a ...

...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute...value that helps you determine whether or not to proceed with a used or new item. To further illustrate ...we mentioned previously.  There is value in the process of creating a budget; you can share your budget...

...presumptive mechanism of action are presented in Figure 2. dCas9 enables sequence replacement without cutting...Nature biotechnology (2016). PubMed PMID: 26789497.  2. Lin, Steven, et al. "Enhanced homology-directed human... a biological process in detail often suggests new strategies to manipulate that process. We began our...

...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore...nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y,...following video to learn Daniel and Matthew's procedures. The video details how to transfect CRISPR plasmids...

...Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication ...Levskaya A, Voigt CA. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010...fact, precise light-based control of biological processes is highly useful in other fields, including synthetic...to directly activate and inactivate biological processes. Chemical effectors are suboptimal for a number...intensity and wavelength can be used to study dynamic processes. Conversely, varied input patterns may also be...

.... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations.   Figure 2: Compensation example based on experimental sample..., you will see the populations move, as in Figure 2, following compensation. Figure 3: Flow plot...In flow cytometry, compensation is the process of correcting spillover from one fluorescent channel to...

...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...sequencing to verify the insert sequence before proceeding with your experiment. Shorter amplicons tend...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...Here, NLuc internally excites the fluor through a process known as bioluminescent resonance energy transfer...

... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W...SSC-A, so most people use that setting. Figure 2: As the cell passes through the laser beam and causes...scale in proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line...heterogeneous cell population, for example from a processed piece of tissue, where cells are present that ...structural complexity. After several steps of tissue processing prior to running my flow analysis, I can however...interest). Let’s double-click inside our gate and proceed to the next plot! Pro tip! If you only get very...

...guides that had zero mismatch off-targets. Guides with 2-bp mismatch off-targets were only toxic if they had...tolerated than mismatches more distal to the PAM, and 2) guides with high GC content have greater off-target...PMID: 28474669. Pubmed Central PMCID: PMC5424143. 2. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide...for identifying genes involved in a biological process. These screens provide an alternative to genome-wide...

...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...dark fusions is likely misfolded, blocked from proceeding through the secretory pathway, and retained in...

...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD... the nick site, which was expected since the processivity window of PolI is 15-20 bp. EvolvR’s on-target...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing...

...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an 8... together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If not...easier with more perspectives contributing to the process Senior people (of which there are never enough,...during the meeting.  Scientists do better with process, so don’t just get together and chat. Have an agenda...started. Have regular meetings. Experiment with the process. Have each person write a Development Plan. Set...