...stimulation itself, the output of the laser must be controlled. As discussed above, each opsin variant has slightly...the A.M.P.I Master-9 Pulse Stimulator, which can control output of several lasers simultaneously, can be.... Pulse generation (TTL pulse) is a standard electronics protocol not specific to lasers or optogenetics...powerful alternatives to the Master-9 that only control one or two lasers are also available. Optimal stimulation...loss of laser power. More recently, wirelessly-controlled optogenetic implants have been reported by several...References 1. Gutierrez DV, et al. Optogenetic control of motor coordination by Gi/o protein-coupled vertebrate... Kim TI, et al. Injectable, cellular-scale optoelectronics with applications for wireless optogenetics...

...do not let the panelists introduce themselves. I do a 1-2 minute introduction. If you chose good panelists...and will play a different role so her ability to control the event is what is important. Choose 4 panelists...logistics To make a really lively discussion, I strongly suggest asking the panelists not to use any slides...it (lapel mics are best). The moderator takes control It is the moderator's job to prevent anyone from...talking, so they will easily take 5 minutes to introduce themselves and, voila, 1/2 of your time is gone...publicity. Good panels do not waste time on long introductions. The moderator asks questions, calling on 1-...

...needed and when is it not? What are the proper controls needed to pressure-test your FRET assay to make...pyrrolysyl-tRNA synthetase Mammalian Cells pCMV-DnpK dinitrophenyl hapten Mammalian Cells pANAP AnapRS Mammalian...commercially available as Lumio Green and Lumio Red from Invitrogen (now ThermoFisher Scientific). These reagents...molecular distances (see Table). However, proper control experiments are needed to establish the dynamic...These experiments include producing a positive control construct where the donor and the acceptor fluorophore... each other on the same protein and a negative control experiment conducted on donor-only labeled protein...biarsenical− tetracysteine motif determined by NMR spectroscopy." Journal of the American Chemical Society 131.13...

... of plasmids, they are generally electroporated into electrocompetent cells to ensure the highest possible...chemical transformations is less than that of electroporation, but an advantage is that chemical transformations...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...DNA in 100 μL of cells over four individual electroporations. The specifics of this can be based on the...5,000,000 30 bp At least 200 ng Liquid Electroporation Protocol (DOCX, 15.8 KB) Barcode SPLINTR... CRISPR Preparing Pooled Libraries NGS Quality Control for Pooled Libraries Resources on Addgene.org Addgene's...

...SARS-COV-1 plasmids. Export control license process    An export control license, once approved, allows...repository, including rabies plasmids, are export-controlled materials in the USA, where Addgene is located...applicable research project when purchasing export-controlled materials. Addgene will then apply for the license...   Due to recent regulation changes, an export control license for rabies materials may be required for...application process.  More information about export control licenses is available through the US Bureau of ...

... they then confirmed by PCR. Subsequent mass spectroscopy studies allowed for the identification of proteins...Fujii Lab generated a transgenic mouse line and retroviral expression system for using this approach in ...analyzed, the Fujii lab also developed another "in vitro" form. In this form, the CRISPR complex, consisting...biotinylated gRNA, were successfully used in this "in vitro" form. Any tag system can be used to isolate genomic...expressed can cause facioscapulohumeral muscular dystrophy (FSHD), a disease for which there is currently... isolation of DNA fragments and chromatin by in vitro enCh IP technology using recombinant CRISPR ribonucleoproteins...as an RNA-guided platform for sequence-specific control of gene expression." Cell 152.5 (2013): 1173-1183...

...Using a spectrophotometer is among the faster methods of quantification. Many spectrophotometers have an... you have multiple contaminating bands or a few strong ones, consider increasing the number of washes ... period of two weeks. After this time, quality control assays showed that the majority of antibodies are...references More resources on the Addgene blog Introduction to Antibodies Plasmid-based Recombinant Antibodies...Protein aggregation - Mechanisms, detection, and control. Int J Pharm. 2018 Oct 25;550(1-2):251-268. doi.../j.cocis.2014.10.002. Randolph TW, Schiltz E, Sederstrom D, Steinmann D, Mozziconacci O, Schöneich C, ...

...for this task. However, most lesioning methods destroy areas larger than intended and may unintentionally...to the neuronal networks involved in directly controlling muscle activity and coordinating movement, several...optogenetics to support these existing findings by controlling neurons in the mPFC with light. Activation of...inhibited by fear/anxiety. On the other hand, the control group preferred staying in the dark compartment...time the animal spends in the passive mode, the stronger is the depressive component of the animal behavior... such genetic study focuses on brain derived neurotrophic factor (BDNF). BDNF is a small protein involved...the same paper, the authors used lentivirus to introduce an shRNA to downregulate expression of miR124a...

... to introduce multiple genetic modifications in the same strain. MazF expression is under control of the... a GFP cassette, and introduced site-directed mutations, all without introducing unwanted selection markers...delivery vector can be integrated into the genome to introduce the genetic modification(s) contained within it...EcoRI and Bsp6I in Escherichia coli in 1995 when introduction of restriction-modification systems onto plasmids... able to monitor lacZ activity over 48 hours in vitro without antibiotic selection. In a subcutaneous ...

...derived from the control trace file. On the plot, the green (edited sample) and orange (control sample) lines...cells, genomic DNA from both edited and unedited (control) populations is PCR-amplified and Sanger sequenced...determined by comparing the edited trace to the control trace. In the ICE algorithm, potential editing ...summary table. You can also use your browser’s Control+F or Command+F functionality to search for a particular...disagreement between the non-edited wild type (control) and the edited sample in the region around the...difference. Traces The Traces tab shows the edited and control, non-edited Sanger traces in the region around ...

...different scientific fields, and knowledge of how to troubleshoot problems at the bench. Specifically, roles in...loads of questions, and I also had to do some troubleshooting as advised by the technical support person....specificity of this antibody?” You will also help troubleshoot products, give general experimental guidance...the world easier. You should also demonstrate a strong willingness to help others with their problems,... initial contact, but is less suitable for troubleshooting or issuing replacement products. In order to...careers are open to experienced professionals with strong interest in the technical aspects of science and...work includes reviewing customer data sets, troubleshooting protocols, analyzing information, and solving...

...unproductive conversations with internet trolls (urban dictionary for trolling: The art of deliberately, cleverly...to write our AMA submission to avoid rousing the trolls. 3. The AMA process was simple and well organized...while it can be anonymous, I can always block the trolls. It seemed like I would really have to open myself...moderators were on top of any comments that were at all trollish. Kudos to the moderators for keeping the quality...discourse. They even advised me on how to alter my intro to reduce the number of potentially hostile posters... the term “implicit bias” which is apparently a troll trigger phrase these days). Redditors ask serious...

...approach complete with freely available positive controls. Routinely test your cell line for mycoplasma ...freezing virus in a dry ice/ethanol bath or liquid nitrogen prior to storing. Know your target cells Labs ...These binding events can be impaired by the electrostatic repulsion of the negatively charged cell with..., A., Garnier, A. “Downstream processing of oncoretroviral and lentiviral gene therapy vectors.” Biotechnology...conditions on the production of lentiviral and retroviral vectors.” Human Gene Therapy Clinical Development...Cronin, J., Zhang, X.Y., Reiser, J. “Altering the tropism of lentiviral vectors through pseudotyping.” Current...Cleghorn G., West M.J., Wei MQ. “The significance of controlled conditions in lentiviral vector titration and...

...Optogenetics Guide γ-Retrovirus Guide Protocols Agarose Gel Electrophoresis Bacterial Transformation... direct shipments weren’t able to clear import control. Partnering with a distributor allowed us to share... How to Design Primers for PCR Agarose Gel Electrophoresis Inoculating Liquid Bacterial Cultures Restriction...Restriction Digest Analysis We also introduced guides — practical primers to help users get started ...we launched the Addgene YouTube channel to add a strong visual component to our educational resources. ... systems to allow for easier access to export-controlled items by streamlining permissions and paperwork...

...field of view. Tetbow can be introduced in vivo either by electroporation of plasmids, or using AAV vectors...type of cell labeling technique involves the introduction of a transgene construct containing fluorescent...cells, the expression level of these XFP is not strong enough for all applications. To resolve this, similar...use in Drosophila melanogaster and is under the control of Flp recombinase. The XFPs in Flybow are membrane-tethered...entire organism or a specific tissue. XFPs are introduced using a plasmid construct called Zebrabow-GateDest...Additional resources on the Addgene blog Read an introduction to live cell imaging Learn more about the origin...

...Stbl4   Electrocompetent cells for cloning and storage of unstable DNA and lentiviral/retroviral vectors...Allows uptake of large plasmids hee   High electroporation efficiency supE44 (glnV44)   Suppression ...cloning and storage, blue/white screening, leucine auxotroph. F- endA1 recA1 galE15 galK16 nupG rpsL ΔlacX74...For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated sequences with...

...system along with precisely truncated attP sites to control recombination of the gene construct. A predictable...gene. Effector genes, any gene that you want to control the expression of using the SPARC system. The effector...rest of the SPARC components and expression is controlled by the upstream recombination outcome. Find ...Find SPARC plasmids at Addgene To more precisely control the amount of labeling, you can alter the attP site...effector position, as well as to optogenetically control the depolarization of R2 neurons using CsChrimson...genome engineering in flies Learn more about controlling gene expression in flies Resources on Addgene.org...

...scientists and plasmid lovers - serves as an introduction to plasmids. Plasmids 101 will provide you with...plasmids contain an origin of replication (which controls the host range and copy number of the plasmid)...the lab are usually artificial and designed to introduce foreign DNA into another cell. Minimally, lab-... that scientists can purify and study it in a controlled setting. Read more: Plasmids 101: Protein Tags...environment Produce enzymes that will make specific, controlled changes to an organism’s genome (genome engineering...guide also contains multiple protocols and troubleshooting tips to make plasmid usage as simple and straightforward...

...mimic aspects of organ morphology and physiology in vitro and have emerged as a powerful model for understanding...for doing gene targeting, reporter generation, controllable gene knockouts, and inducible gene inactivation...design. B) Strategy for leak-free transcriptional control. C) Iterative transduction workflow overview. Image...SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore (circularly permuted...the cpGFP without altering that of the internal control (Fig 1). Because this method relies on the fluorescence...-500nM range) and a high specificity for SLs in vitro, and they further demonstrated that when tested ...

...activity in the brain was measured with implanted electrodes. The Archon1 voltage reporter allows a way to...cre-dependent version at Addgene! Fluorescent protein and control AAV These fluorescent protein expression vectors...interneurons. These vectors can also be used as the control vectors for experiments encoding a fluorescent-... Addgene in the next 6 months, pending quality control. For an estimate on availability date or to be ... and serotype you’d like to be notified about. Controls Plasmid Serotype Name 114472  AAV5, AAV8 ...Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST Chemogenetics Plasmid Serotype Name ...