...fluorescent signal. Sanger can sequence approximately 500-1000 bases downstream of the known primer region with...isolating one specific DNA sequence. This obstacle has been overcome by the field of recombinant DNA technology...isolates, the K-12 strain and the B strain. K-12 has led to the common lab strains MG1655 and its derivatives...polymerase. Thus to replicate a piece of DNA in vitro one has to know some of its sequence to design a effective...Epitope Tags Tag Amino Acid Sequence FLAG DYKDDDDK HA YPYDVPDYA His HHHHHH Myc EQKLISEEDL V5 GKPIPNPLLGLDST... dissolved in dH 2 0. Addgene recommends making 1000X stock solutions and storing aliquots at -20°C. To...media, add 100 μL of a 100 mg/mL ampicillin stock (1000X stock) to 100 mL of LB. DNA sequencing for plasmid...

...these adaptive bacterial and archaeal immune systems has led to extreme sequence divergence that requires ...landmark publication in 2013. Since that point, it has become the most engineered Cas enzyme, with adaptations... CRISPR type that exclusively targets RNA. Cas13 has become a natural successor to traditional RNA interference...and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013 Additional resources on ...

...effective than both AAV9 and AAV5, which has tropism for lung cells, and has increased overall transduction throughout...this definition: A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...both). Therefore, a high-efficiency systemic capsid has better efficacy than AAV9 and/or its intermediate...used in LY6A-nonpermissive mouse strains. **AAV1-X1 has not been tested in rats. For more information on ...neurons in the CNS (Chan et al., 2017). AAV-PHP.eB has frequently been used for screening candidate regulatory...depositing manuscript) across the brain. AAV-BI-28 has been tested in C57BL/6J mice and effectively transduces.... 4c, Jang et al., 2023; Brown et al., 2021.) It has a similar transduction rate to AAV-PHP.eB in the ...

...Research will be held from 22-24 October. This blog has been contributed by guest blogger and Future of Research... of a system that, despite all of its successes, has many deep-rooted flaws (1). As we discovered through...example, Harvard’s Therapeutics Graduate Program has a compulsory internship to be taken in an industry...Marder, professor of Biology at Brandeis University, has recently published an article arguing against reducing...Labor and Worklife Program at Harvard Law School. He has continually promoted the cause of increasing data...Research: a perspective from junior scientists." F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central...

...CRISPR/Cas9 technology has been a breakthrough for the genome-editing field. It has the possibility to treat... applications.” Li’s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of... expanded organoids. “CRISPR in adult stem cells has huge potential, because the genomes of adult stem...& Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6. Wu et al. 2020. . “Advances in CRISPR/Cas-based...

...sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process may... immune response to it. Once the immune response has developed, the antibodies are collected, tested against...epitopes on constant regions of other antibodies. This has useful research applications, and is covered in the...exist as monomers. Each antibody class therefore has a valency, or number of binding sites available per...of antibodies multiplied by two, as each antibody has two binding sites. Isotype Description Configuration... must be able to detect if and where an antibody has bound. Signaling molecules , proteins that emit a...to avoid confusing data. Check that your machine has appropriate lasers and filters for each fluorophore...

...shRNA sequences pLKO (Plasmid #10878) This plasmid has been used by The RNAi Consortium, also known as the...Plasmid Kit Retroviral Barcoding Library (Kit # 1000000097) The Winslow lab barcoding library consists of...Multiple Lentiviral Expression System Kit (Kit #1000000060) The MuLE (Multiple Lentiviral Expression) system...

... in situ. Used since the mid-90s, this technique has revolutionised the way we apprehend molecular complexes...protein interactions in living cells. Since then FRET has also been used to measure conformational changes,...brightness - even though in the most popular pair, CFP has five-fold less brightness than YFP. The stoichiometry...Fluorescence lifetime imaging microscopy method FLIM has been developed more recently and is the most rigorous...emergence of faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...Bioessays. 2012 May; 34(5):341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance...

...Adeno-associated virus (AAV) has many features which make it a great viral vector, but its packaging ... Overlapping  Overview: The overlapping strategy has a region of homology, typically ~400-1400 bp long...Drawbacks:  Generally an overlapping split vector has equal or higher transgene expression when compared...split AAV method While the hybrid dual vector system has a slight advantage over the overlapping and trans-splicing...efficiency of transgene expression from split vectors has been inconsistent. A greater understanding of basic...al., 2002 used an overlapping vector with 440 or 1000bp overlaps to deliver the alkaline phosphatase gene...

...kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which has the N-terminus domain...proteins in minutes rather than hours like BioID. APEX has allowed scientists to identify the proteomic map ...BioID was developed in 2012, proximity labelling has become a powerful technique for studying protein-...proximity-based labeling methods for interactome mapping. F1000Research, 8.  ...

...trimers, tetramers). Since then, multimerization has been observed to occur in many different kinds of...sequences. In some specific cases, multimerization has been observed to occur more frequently in plasmids... Ausubel, 1976). In fact, if your plasmid sample has a mix of different-sized multimers, you may even ...~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...multimers is capillary gel electrophoresis (CGE), which has been used in plasmid vaccine and gene therapy development...Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/elps.201000061. PMID: 20589859. Levene SD. Analysis of DNA topoisomers...

... cell mobility within these complex environments has never been easy and, for many years, researchers ...Intravital imaging powered by two-photon microscopy has uncovered host-pathogen interactions leading to the...proteins can be used in your experiments, experience has shown that some are more reliable and more efficient...important to finely tune the excitation wavelength as it has been shown that even a small incremental change of...this system called Brainbow, the Ubow mouse strain has been created to fate map Langerhans cells and follicular...with Toxoplasma gondii." PLoS Pathog 5.7 (2009): e1000505. PubMed PMID: 19578440. PubMed Central PMCID: ...

...invention in the late 1990s, Gateway cloning technology has become very popular as a rapid and highly efficient...attL-containing entry vector.   Pro Tip: Addgene also has ready-made entry clones available for many popular...Drawbacks. Clon Transgen 4:138. doi:10.4172/2168-9849.1000138 2. Hartley JL. Use of the Gateway System for...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...calculate the titer for each sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume...sealer and press the ‘Seal’ button. After the plate has been sealed, proceed to thermocycling. Thermal Cycling...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...polyclonal cell population, meaning that the transgene has integrated in different locations in the various ...variety of dilutions and pick the population that has the most desirable level of expression. Over time...

...reproducible” does not necessarily mean that someone has replicated it precisely. You can find a great discussion...easier to understand. The need for great stories has, however, made it difficult to publish so-called .... Finally there are new publishing outlets like F1000, PeerJ, PLoS, and bioRxiv that make it easier to...

...Future of Research believe that, while the mandate has been removed, institutions should still go ahead ...blogpost and more recently in a paper published in F1000Research on November 2016. Due to the nature of the postdoc...

...middle is hundreds, and the third is tens. Therefore, 1000 µL would read as 100 from top to bottom (as shown... flow rate into the pipette tip. Once the liquid has finished drawing into the pipette tip, lift the pipette...Anatomy of a Pipette This front view photo of a P1000 pipette shows the plunger at the top, which is used...tip off of the pipette. This side view photo of a P1000 pipette shows the volume adjustment ring right below...come in standard sizes: P2, P10, P20, P200, and P1000. The table shows the pipette with the appropriate...This image shows the P2, P10, P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume... 2 to 20 µL P100 10 to 100 µL P200 20 to 200 µL P1000 100 to 1,000 µL Each pipette should be used with...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...mL) x dilution factor e.g., If the 1:100,000 well has 75 colonies, then there are 75 colonies /1.5 mL x...

...pipettes Ice bucket to hold antibiotic Antibiotic at 1000 x concentration disolved in the appropriate liquid...darken during the autoclave process if your sample has spent at least 10 min at 121 ℃. Use lab tape to label...antibiotic to the molten gel mix, you should create a 1000x stock solution. For example: If you’ll be preparing...