...genetic underpinnings of these processes has been complicated by the lack of precise genetic tools for modulating...recent advancements and offer commentary on their application to neuroscience research. What is CRISPR-based...accessible to neuroscientists, not all neuroscience applications require such optimized tools. This is elegantly...necessitates such molecular gymnastics for sensitive applications such as CRISPR screening. The plasmids used ...used by the Qi Lab for this particular application can be found here.  Why CRISPRa? Although cDNA overexpression...gene activation in select in vitro and in vivo applications, the tools described above make CRISPRa in the...relatively easy and accessible. There are also other applications for which CRISPRa is well-suited, including ...

... not need to be run in triplicates. ddPCR is an extremely accurate application, even in areas of the genome...blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative/real-time...standard curves, and normal controls must be run in triplicate.  The nice thing about qPCR is that since it ...fluorescent probe and quencher to quantify specific amplicons.  Sybr has high background noise, due to the large... affected by AT-rich DNA and the length of the amplicon and it has a higher rate of false positives (Colborn...is not affected by AT content or length of the amplicon. As the probes can have several different markers...range of RQ values as noted above.   In both applications, results can be affected by DNA quality and ...

...few principles that will help you resolve most conflicts, even with difficult people. If I had to use one...strategies to become more assertive & resolve conflicts What Is Assertiveness? An assertive person is ... side-tracked, and you might create even more conflicts. 4) Define in advance how you would like the problem...in writing One of the most frequent sources of conflict is miscommunication. For example, you might misunderstand...you to resolve both personal and professional conflicts. At the workplace, these strategies will help ...miscommunication, which can lead to even more conflicts. If your supervisor is a difficult person, the...an inexperienced manager, you can resolve most conflicts if you communicate your ideas assertively, listen...

...species or metabolic pathways. A new study takes us one step closer to the vision of metabolic biofactories...s vision of ‘biofactories’, most successes in metabolic engineering have been limited to the overexpression...be host-independent, beyond the oft published metabolic engineering realm of S. cerevisiae and E. coli...control via scRNA design is readily apparent in metabolic engineering. The authors elegantly demonstrated...demonstrated the potential of this to control a metabolic network in S. cerevisiae by using three RNA programs ...programs, the authors sequentially rerouted the metabolic flux to generate differently pigmented derivatives...regulatory switch. By no means limited to the field of metabolic engineering, this is such a powerful and flexible...

...Sharing Policy Discussion PLOS’ New Data Policy: Public Access to Data PLOS’ New Data Policy: Part Two...supporting the research reported in PLOS publications. The policy was not an overnight revelation, rather...advice about the scope of the policy. End of story? Not quite. The PLOS policy on data access is a formal...Data Sharing Policies, etc.) and is founded on the principle that scientific publications must contain...clarified what the policy intent was and provided an FAQ list to further explain the policy. The fact that...This post was contributed by Jim Woodgett. Public Library of Science (PLOS) created a stir earlier this...year when it announced its data access and sharing policy. Since early March, the open access publisher has...

... : Kwabena's public health research, the HEAD outreach program, and educating the public about dietary...awarded plasmids from Addgene as a successful applicant to the Addgene, Seeding Labs Plasmid Grant. For...bring new opportunities to Ghanaian scientists Click here to listen! Podcast breakdown (Time : Topic)... public health...

...Initiative. Endy's team made news in March with a publication in Science describing the development of “transcriptors...are now available at Addgene through the Biobrick Public Agreement (BPA) Plasmid Collection along with a...It's wonderful to have Addgene as a professional public-benefit partner who shares our core values." Addgene...BIOFAB and BIL gates plasmid kits, the Biobricks Public Agreement, and the future of the bioeconomy. Addgene...work, and it begins with the central dogma - replication of DNA, transcription of DNA to make RNA, translation...

...research. This year, we put out another call for applications. We had an overwhelming response and were excited...sensor with proof-of-principle experiments that replicate current knowledge, but the sensor can also kinetically...  Michal Bernach: Developing an artificial leaf replica system to study plant-bacteria and bacteria-bacteria..., Bernach sought to develop an artificial leaf replica system. “This bottom-up approach will allow me ...Bernach explains. Developing an artificial leaf replica means creating a “leaf” with similar permeability...fluorescent proteins are made. As the bacterial replicate, fluorescent proteins will be diluted out and ...

... bacterial expression system such as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant...bacteria. Learn more about the different CRISPR applications in plants.     Transgenic yeast as source of...

...bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope tag (6xHis...types of yeast expression vectors. Consider your applications and experimental design. You can create a new...more traditional approach of an autonomously replicating vector that carries either a wild-type version...a variety of other types of vectors for most applications used to study gene function. From making transgenic...

...(Lenti_sgRNA_EFS_GFP) 65656 Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA(F+E) 59986 Other...RNA Targeting RNA Targeting RNA Editing Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled...

... all publications arising from the use of this vector. A.2 Map of pLKO.1 pLKO.1 is a replication-incompetent...terminal repeat. f1 ori f1 bacterial origin of replication. Amp R Ampicillin resistance gene for selection...bacterial cells pUC ori pUC bacterial origin of replication. 5’LTR 5’ long terminal repeat. RRE Rev response...it has been diluted. Mix by swirling or gently flicking the tube. Incubate for 5 minutes at room temperature...the walls of the tube. Mix by swirling or gently flicking the tube. f. Incubate for 20-30 minutes at room... on the size of your target plate. TIP: MOI (multiplicity of infection) refers to the number of infecting...Appendix I.1. Sequence of pLKO.1 TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning vector...

...) - LiCl selectively precipitates RNA versus DNA or proteins. Add the correct amount of 7.5 M LiCl solution...solution to bring the concentration of LiCl in the extracted aqueous layer to 2.5 M. Incubate at -20°C ... not affect the quality of RNA or downstream applications. *Pro-Tip* To improve yield of RNA, instead ...

...Load 5 μL of each standard in duplicate Load 5 μL of each sample in duplicate. Do not forget to include a...dilution is appropriate). Quality of duplicates: Exclude duplicates from analyses if there is more than...Considerations Always run standards and samples in duplicate at least If possible, include an AAV reference...standard with DNase ** Make 6 serial dilutions, in duplicate, of your standard curve plasmid (2x10 9 stock ...

...The amount of DNA that you cut depends on your application. A diagnostic digest typically involves ∼500 ...volume usually varies from 10-50 µL depending on application and is largely determined by the volume of DNA...manufacturer’s instructions. *Pro-Tip* Depending on the application and the amount of DNA in the reaction, incubation...you will be using the digested DNA for another application (such as a digestion with another enzyme in a...

...bacteria are used as the means for both storing and replicating plasmids. Because of this, nearly all plasmids...expression) carry both a bacterial origin of replication and an antibiotic resistance gene for use as ... microcentrifuge or falcon tube. GENTLY mix by flicking the bottom of the tube with your finger a few ...

...antibody can be purified for use in a variety of applications. Sharing speeds science. We believe that sharing...antibody-expressing plasmids containing the SV40 origin of replication use a HEK293 line stably expressing the SV40 ... containing the Epstein-Barr virus origin of replication use a HEK293 line stable expressing Epstein Barr...

...sufficient quantities of library for experimental applications. Repeated amplifications should be avoided as...required for bacterial propagation (origin of replication and antibiotic selection). This recombination...each 50 µL aliquot of thawed Endura Duos on ice. Flick gently to mix. Electroporate cells (one at a time...

... lentiviral vector allows you to control the multiplicity of infection (MOI) in downstream studies. This... these cells can still be used in downstream applications. Safety Warnings Lentivirus is generally considered...

...lentivirus can be used for a variety of downstream applications such as stable-cell line generation. Last Upload...DNA. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15-...