...of experimental results in the order they were performed does not make for a memorable presentation anywhere... just plan to reuse a prepared talk unless it perfectly fits the occasion.  I speak on the same topics...

..., or knocking down YGOI through RNA-mediated interference (e.g. L4440) and observing the downstream effects...additional details and guidance for choosing your perfect plasmid backbone. Or just ask –  leave a message...

...themselves.  Here is my experience looking for the perfect job. Summer work experience in cancer research... good job to list on my CV. Three months was a perfect duration for me because it was sufficient to figure...University of Queensland in Australia, this was a perfect opportunity to work in a genetics lab. As a UK ...acidemia to have the best chance of finding their perfect job.   Resources at Addgene Read Additional Career...

...microenvironments. These screens have primarily been performed in activated T cells, precluding analyses of T...cells. More broadly, this system can be used to perform in vivo screens in other cell types. A key feature...in vivo screens. Figure 1: Using FITS to perform in vivo CRISPR screens to assess T cell responses...

...that function downstream of DNA, such as RNA interference, can reversibly exert their effect independent...independent of gene copy number. RNA interference (RNAi) is a conserved eukaryotic process where approximately...DCL) process the dsRNA into specific types of interfering RNAs.   RNAi vs. CRISPR: Complementary techniques...from a long hpRNA trigger, only a single ~21-nt interfering RNA is produced using shRNA/amiRNA. Although ...Hiwatashi Y, Goshima G (2012) An Inducible RNA Interference System in Physcomitrella patens Reveals a Dominant...Doudna JA (2013) Molecular Mechanisms of RNA Interference. Annu Rev Biophys 42:217–239 . https://doi.org...

...developed in 2012, proximity labelling has become a powerful technique for studying protein-protein interactions...

...Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922. PLoS ONE 11:e0154027 . https:/...

...function by RNA interference (RNAi) RNA interference (RNAi) is a powerful tool for performing targeted knockdowns...the students performing these experiments have a good baseline for comparison. Performing WISH using these...are too few resources to teach students how to perform these experiments in the beginning. In most classrooms...observation and dissections. Students don’t usually perform more modern techniques used in genetics or molecular...addition, the Sánchez lab has made it easier to perform advanced developmental biology experiments in planarians...genes. Double-stranded RNA (dsRNA)-mediated RNA interference (RNAi) to knockdown expression of specific gene...both WISH and RNAi experiments, as well as how to perform worm amputations. They also created an online companion...

...than one another, a two-tailed test should be performed. This test will assess if the mean of an experimental...tests, referred to as post-hoc analysis, must be performed. These will be discussed later. One and two-way...within your ANOVA/ANCOVA analysis. If you have performed a MANOVA/MANCOVA and want this information, multiple...multiple ANOVAs/ANCOVASs must be performed first, and then post hoc tests can be run on these tests. ...can reduce the family-wise error rate by only performing the comparisons you really care about. Family-wise... positive occurring when many comparisons are performed. For example, if your alpha is set at .05, then...test will be incorrect. If you have 5 samples and perform every pairwise comparison (10 in total), your likelihood...

...optimization as each antibody performs differently and some antibodies impact the performance of other antibodies...beautiful results. Performing an immunofluorescence experiment The process to perform IF can be broadly...important considerations to make when planning and performing an IF experiment. What is immunofluorescence... different species and colors makes it easy to perform multiplexed IF. By using multiple primary antibodies...in IF experiments. Precisely which controls you perform will depend on your experimental context.  Choosing... to use, then the primary antibody will likely perform as expected in both.  How do you find validated...may be, it is always important to verify their performance in your hands and in your specific experiment...

...pooled libraries have allowed scientists to easily perform genome-wide screens to effectively and efficiently...human genome-wide CRISPRko (knock out), CRISPRi (interference), and CRISRPa (activation) libraries that do...that improved sgRNA activity and thus library performance. The Root and Doench lab’s optimized genome-wide...CRISPRi, and CRISPRa libraries have been shown to outperform other popular CRISPR libraries (see below). Let...colon cancer) cells and assessed the libraries performance via the depletion of a gold standard set of genes...developed sgRNA-level metrics to assess library performance in viability screens. Based on these metrics ...sgRNAs, Brunello even with only a single sgRNA outperformed another CRISPR library with 6 sgRNAS, highlighting...

...protocols and performing rigorous quality control (QC). The specific QC experiments performed varies for ...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...and stored at -80 °C. Titer Titering is either performed by Addgene or by the University of Pennsylvania...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....Pennsylvania Vector Core is (as of April 2016) also performed by ddPCR.  Quality Control Addgene ensures high.... To learn which specific QC experiments were performed on your lot, please contact us. Full sequencing...the Viral Genome Next-generation sequencing is performed on viral genomes isolated from the final AAV preparation...

...your antibody performs as expected in one application does not mean that it will perform as expected in...doesn’t necessarily mean that you yourself have to perform extensive validation experiments for every antibody...data, you may need to roll up your sleeves and perform some extra validation experiments yourself.  Goal...researchers can use to show that their antibody performs as expected. One of the benefits of these pillars...levels tend to correspond to protein levels, so you perform immunohistochemistry with your antibody on these... the epitope recognized by your antibody. You perform immunocytochemistry on these mutant cells as well...caveat is that different antibodies may or may not perform well under the same conditions. Example: You want...

...homemade competent cells of 107 CFU/μg are perfectly fine, yet performing multiple modifications will require... pathway to allow any cloning procedure to be performed using a simple two-step, 2 hr protocol prior to...adopted by any laboratory. All procedures are performed using the same three step protocol:  Fragment...efficiency of the strain: when asking the cell to perform multiple recombination events, all linear fragments... design (Figure 2), IVA cloning can be used to perform any plasmid modification, from insertions, deletions... reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride the old codon, amplifying...primers can be used in combination to simultaneously perform modifications, simply by adding all the necessary...

... line to perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments...CRISPR-Cas system localizes to unintended sites and performs its programmed function there, this is an off-...off-target effects, but it’s important to not let ‘perfection’ get in the way of ‘good enough’. Predicting ...would confound your experimental results and 2) perform targeted off-target measurements at those sites... to quantify off-target effects entirely is to perform whole genome sequencing. While this may be unnecessary... The con to these approaches is that NHEJ must perform the repair to allow for detection, not Homologous...targets as well as clonal differences. If you are performing CRISPR library screens, the same principle holds...

...contrast technique in modern biology (1). When performed correctly, fluorescence microscopy provides a ...contrast image in which a bright signal overlays a perfectly black background. In addition to this, the use...confocal microscopes use parallelization to enhance performance. In spinning disk microscopy, a metal disk containing...coverslip at an angle such that when it reaches the interface between the glass coverslip and a sample in aqueous...within a few hundred nanometers of the glass/water interface. Super resolution microscopy Super Resolution ...means that the images will appear sharp without interference from blurry out-of-focus light. The most difficult...be needed. Most commonly these experiments are performed on a fluorescent widefield microscope. Here, excitation...

...extracellular fluid into the perfused liquid. The probe then collects the perfusion liquid for quantitation...fluid (Darvesh et al., 2011). This technique is performed with a probe surrounded by a semipermeable membrane... quantitation is desired (Figure 1). The probe perfuses liquid, which causes the passive diffusion of ...The advantage of this method is that it can be performed in a live awake animal by surgical implantation...into a specific brain region. While the method is powerful and offers immediate measurements in a live animal... GRABDA sensor here! Overall these sensors are powerful new developments in the detection and measurement...

...predict how fluorophores perform in ‘real experiments’, it is advisable to perform time-lapse imaging of ...To determine photostability, an experiment is performed that measures fluorescence intensity over time...Therefore, photostability measurements can be performed with soluble fluorescent proteins or localized...photostability measurements reported in the literature are performed in different ways. We describe a number of issues...fluorescent protein. Photostability measurements can be performed on purified fluorescent proteins. To avoid diffusion...changes in excitation power, it is important to perform the experiments on a single day on a stable system...

...donated MiSeq.  For Complete Plasmid Sequencing we perform a 2x251 run on our MiSeq that takes about two days...involves a significant amount of QC. First, we perform Picogreen quantification on each and every isolated...samples for Viral Genome Sequencing (VGS). QC is performed throughout the library preparation process and...Aligning the sequence The first step we take when performing quality control of newly deposited plasmids is...backbone sequence. We’re not necessarily expecting a perfect match - we will often find a few mismatches in ... reads to the correct sample. The PCR is then performed, optimized as needed, and cleaned up using a standard...to remove any template DNA and primers.  After performing extensive QC on that sample, we’ll then load ...

...for endogenous gene activation Optogenetics is a powerful tool that utilizes light to control and monitor... affinity for SspB1. In order to create a more powerful LID, the authors recently used computational protein...pathways. These LIDs are generalizable, versatile and powerful tools which allow you to easily and reversibly...elements in a single lentiviral vector provides a powerful tool to modify mammalian cells. Multicolor CRISPR...and broad host-tropism. These tools open up many powerful avenues through which one can examine molecular...all-optical electrophysiology is designing the perfect voltage indicator and optogenetic actuator pair...expression via western blot, to purify your POI, to perform pull down experiments like co-IPs and ChIP, and...