We narrowed to 731 results for: Sis

...homology, allowing the strands to anneal. DNA synthesis then fills in the gaps. The end result is a deletion...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus...into the FBL locus. Upon sequencing of puromycin resistant clones, they found that 80% and 50% of clones ...References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with...and Ling Lu. (2016). “Highly efficient CRISPR mutagenesis by microhomology-mediated end joining in Aspergillus...

...namely that they can cause cancer via insertional mutagenesis (which HIV does not do). Furthermore, molecular... thanks to Addgenie Rebecca Hutcheson for her assistance with lentiviral replication.  Resources and ...1974). Infectious rous sarcoma virus and reticuloendotheliosis virus DNAs. Journal of Virology, 14(5), ...polymerase of Rous sarcoma virus: Initiation of synthesis with 70 S viral RNA as template. Journal of Molecular...Full-Length Human Immunodeficiency Virus Type 1 DNA Synthesis. Journal of Virology, 82(5), 2376. https://doi.org...

...human CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per...depletion of essential genes. In the subsampling analysis of the sgRNAs, Brunello even with only a single... a comparison study screening for vemurafenib resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic...Mediator) uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA...reading frame (ORF) overexpression libraries for resistance to MEK inhibition. Although both Calabrese and...

...recombination.    Designing donors for HDR The basis of the HDR pathway relies on repair from a template...molecule, which is usually an endogenously available sister chromatid, though exogenous DNA can be used as ...kb), a dsDNA donor will need to be used due to synthesis limits of oligonucleotides. These donors traditionally...predominantly in S and G2 phase of the cell cycle, when a sister chromatid is likely to exist as a repair template...long. Prolonged cell cycle arrest can lead to apoptosis in as little as 10 hours, so read up on the timing...

...The ACE reporter is a bicistronic construct that consists of a mutated mCherry and a downstream constitutively...mScarlet reporter, luciferase reporter, and neomycin resistance reporter. Image adapted from Katti et al., 2020...As a proof of concept the Dow lab tested their hypothesis with GFP fluorescence. To do this they generated... including mScarlet-I, luciferase, antibiotic resistance markers, and other enzymes such as Cre-recombinase...proteins including fluorescent proteins, antibiotic resistance, and luciferase. References Gaudelli NM, Komor...

...yourself using PCR, but gene synthesis can save you time. Using gene synthesis, you can request the sites...before you start doing your cloning. Most sequence analysis software includes Golden Gate cloning tools that...lot of headaches down the line! Cash in on gene synthesis Before you start your cloning, it is advisable...direction (which is not easy to see in colony PCR electrophoresis), is free of undesired mutations, and that ...

...available to confer greater transcriptional control, assist with proper protein folding, and deal with sub-...in outer membrane protease VII, which reduces proteolysis of the expressed recombinant proteins. lon protease...deleted (designated lon or Δlon) similary reduce proteolysis of the expressed proteins. hsdSB (rB- mB-): These...coli B strain, except ** which are K12 Strain Resistance Key Features Genotype Use BL21 (DE3)   Basic.... The pLys plasmid contains a chloramphenicol resistance cassette for positive selection and a p15A origin...

...possible by tools that allow cells to become persistently fluorescent during development. Stochastic multicolor...fluorescent neuron is visible. Chemical tags are more resistant to tissue clearing methods and can be labeled ... "Multicolor fate mapping of Langerhans cell homeostasis." Journal of Experimental Medicine 210.9 (2013...genetic multicolor cell labeling for neural circuit analysis in Drosophila melanogaster." Nature methods 8.3... Snippert, Hugo J., et al. "Intestinal crypt homeostasis results from neutral competition between symmetrically...

...correctness of the hypothesis. The results of experiments can help support or reject a hypothesis. Looking at...Addgene or its staff. In Washington, D.C. as well as sister locations on April 22, 2017, scientists and non-scientists...knowledge. The process begins with a testable hypothesis - an educated guess about how some part of the... Magazine: A grim budget day for U.S. science: analysis and reaction to Trump’s plan Science Magazine:...

...seven-repeat proteins: Sjuts, Hanno, et al. "Molecular basis for inhibition of AcrB multidrug efflux pump by ...system in correlation to the size of the product. Analysis of protein size and expression level can be conveniently...frequencies across timepoints or populations. This kit consists of a 48 plasmid adapter library, with each plasmid... daptomycin reveals an ordered progression to resistance." Antimicrobial agents and chemotherapy 57.11...effects during adaptive evolution of antibiotic resistance." Proceedings of the National Academy of Sciences...

...function is to regulate DNA repair, senescence, and apoptosis. Work by Kamio et al. (2025) utilized HiBiT to...characterize the proteasomal degradation of cystic fibrosis transmembrane conductance regulators (CFTR) (Kamada...ion channel, mutations of which lead to cystic fibrosis, one of the most prevalent inherited diseases.... these challenging proteins. VEGF-A mediates angiogenesis through its principal receptor tyrosine kinase...focuses on commercializing their cell and protein analysis products. You can find plasmids deposited by Promega...

...antibodies as major culprits in the reproducibility crisis of biomedical research (Baker, 2015; Bradbury &...approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from several...protein interactions could be a factor in your analysis. Example: You are planning an RNA-immunoprecipitation...and send the captured proteins off for mass spec analysis (above). When the results come back it shows that...here!  References Baker M (2015) Reproducibility crisis: Blame it on the antibodies. Nature 521:274–276...

... the cause of leaky expression. To test their hypothesis, they randomly scrambled one pair of lox sites...packaging even begins. This spontaneous recombination persists even in recombinase deficient E. coli and under...References Aschauer DF, Kreuz S, Rumpel S (2013) Analysis of Transduction Efficiency, Tropism and Axonal...Ersing I, Haery L, Harten DeMaio K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L, Mueller... (2020) A Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity of Recombinant...

...complementary to the dsRNA (Wilson and Doudna, 2013). RNAi consists of two non-exclusive gene silencing pathways: ...different Dicer-like enzymes are involved in the biogenesis of different small RNAs (Figure 1). These siRNAs...amiRNAs based upon miRNA precursors of rice and Arabidopsis species, respectively. Short hairpin RNA (shRNA...plants, such as in the model flowering plant Arabidopsis thaliana (Li et al., 2013; Zhang et al., 2018...plants outside of limited, single cell protoplast analysis. Instead, microscopic plants must be visually ...

...guest blogger, Kaustubh Kishor Jadhav, a Research Assistant at MGMs Institute of Biosciences and Technology.... The absence of a rigid cell wall makes them resistant to antibiotics and antibacterial drugs like penicillin...of antibiotics can allow mycoplasma to develop resistance to the antibiotics. Antibiotics must only be ...prolonged use of a particular antibiotic can induce resistance for other bacteria apart from mycoplasma. Always...Jadhav! Kaustubh Kishor Jadhav, MSc is a Research Assistant at the MGMs Institute of Biosciences and Technology...

...replace a gene of interest with an antibiotic resistance cassette.   Substrate DNA Design and Generation... selectable DNA fragments, such as antibiotic resistance genes, as well as non-selectable DNA fragments...red phage, λTetR, that carries the tetracycline resistance gene and the lambda red repressor cI857.  Once...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...first be selected via antibiotic resistance, but all clones should be further tested to confirm the presence...

...extra PGC-1a1 only in their skeletal muscle show resistance to stressful circumstances – including unpredictable...Activation of the PGC-1a1 Pathway Gene expression analysis of the animals’ skeletal muscles led to another...mck-PGC-1a1 mice, and those who exercise, show resistance to depression. For the researchers led by Dr...t what they’d expected. "Our initial research hypothesis was that trained muscle would produce a substance...

... sequence similarity, phylogenetic analysis, neighborhood analysis of surrounding genes, and experimental...sequence divergence that requires a multi-pronged analysis to sort effectively. The current classification...are merely linked by a similar function. After analysis, CRISPR classifications were placed into a fairly... of factors, including your goal, experience, hypothesis, and even the time and money available. The breadth...

...FP-chimeric protein may be at any one of many stages of synthesis and degradation. For this reason it is virtually...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...negative” photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't...is blocked during the Bax activation phase of apoptosis." J Cell Biol 164.4 (2004): 493-499. PubMed PMID...PMC1635685. 12.Paez-Segala, Maria G., et al. "Fixation-resistant photoactivatable fluorescent proteins for CLEM...

...higher volume can affect the lysis buffer ratio and result in incomplete lysis. You may be able to scale ..., which can happen if you leave the sample in a lysis buffer for too long or if you get a little overzealous...scale the amount of buffers used to ensure proper lysis, but you will still face column capacity limitations...troubleshooting. If you need a high level of consistency between lots, then reviewing their quality assurance...