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We narrowed to 136 results for: THY-2

Showing: 31 - 40 of 136 results
  1. Plasmids 101: TOPO Cloning

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    Blog Post
    ... the two DNA strands and releasing topoisomerase (2).  These days commercially available TOPO kits provide...Taq polymerase using your favorite PCR protocol.  2. Set Up TOPO Cloning Reaction: Mix together the PCR...PubMed PMID: 1658796. PubMed Central PMCID: PMC52876. 2. Novel approach to molecular cloning and polynucleotide...ability of complementary basepairs adenine (A) and thymine (T) to hybridize and form hydrogen bonds. This ...vectors or cloning arms with overhanging 3´ deoxythymidine (T) residues that are covalently linked to ...
  2. Light Sheet Fluorescence Microscopy

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    Blog Post
    ...Carballosa, et al, (2015) eNeuro, March/April 2015, 2(2) e0001-15.2015. Pubmed. Resources: http://openspim.org...orthogonal-plane fluorescence optical sectioning (OPFOS) (2). Arne Voie, David Burns and Francis Spelman focused...in LSFM. In LSFM the laser light sheet, typically 2-6  microns, illuminates only one thin plane of the...(10).  He used a solution of benzyl alcohol and methyl salicylate which was later modified by others to...subpopulation of neurons are brightly labeled with GFP (e.g. Thy1-YFP-H mice).  However, we recently developed AAV...
  3. Plasmids 101: Knockout/Knock-In Plasmids

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    Blog Post
    ...the addition of Cre recombinase (Figure 2). Figure 2: A knockout targeting vector designed to... through an experiment designed to knock out exon 2 of a given gene.   Figure 1: A knockout targeting...For recombination to occur in a cell, a minimum of 2 kb of sequence homology is required, but 6 to 14 kb...construct to your target cells. After recombination, exon 2 of the targeted gene will be removed from the chromosome...the GFP/NeoR cassette is inserted in place of exon 2. Thus, the targeted gene is disrupted (knocked out...recombination event will remove NeoR only and leave exon 2 floxed, as seen in line 4 of Figure 4. Since the loxP...genome, a negative selection marker like the HSV thymidine kinase (HSV-tk) is included just outside one of...
  4. Illuminating Epigenetics with A FRET Based Biosensor

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    Blog Post
    ... constructed 2 versions of this reporter to target the K9 (pcDNA3-K9 histone methylation reporter) and... 525.7570 (2015): 533-537. PubMed PMID: 26352475. 2. Bygren, Lars O., et al. "Change in paternal grandmothers...in their DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG islands...hybridization, methylation-sensitive restriction enzyme digestion, DNA adenine methyltransferase identification...linker is a methyllysine binding domain (chromodomain), which binds selectively to lysine-methylated peptides...the results. When methylation occurs at the histone-derived peptide, the methyllysine binding domain causes...to recognise lysine 27 methylation. When tested in living cells where methylation of these sites was controlled...
  5. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

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    Blog Post
    ...generate 2 possible products: 1) a partially dsDNA duplex with single-stranded 3’ overhangs or 2) if the...recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required for recombineering...generic lambda red recombineering experiment (Figure 2). In the following sections, key steps that differ...confirmation of recombinant clones.    Figure 2: Overview of using Lambda Red recombineering system...pairs of homology to the targeted insert site (Figure 2, top).  These primers are typically ~70 nucleotides...bacteria that has key MMR proteins knocked out or 2) specially design ssDNA oligos to avoid MMR: 1) E....will have more unintended changes to their genomes. 2) Designing ssDNA oligos that avoid MMR activation:...
  6. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://doi.org/10.1089/hgtb.2013.131 Lock,... virus Once you have transfected your cells, wait 2–5 days and then collect the supernatant and/or producer...quantification of adeno-associated virus serotype 2-derived inverted terminal repeat sequences. Human ..., R., & Kleinschmidt, J. (1999). Titration of AAV-2 particles via a novel capsid ELISA: Packaging of genomes... genomes can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038...Characterization of a recombinant adeno-associated virus type 2 Reference Standard Material. Human Gene Therapy, 21...rAAV from your supernatant, you can simply use polyethylene glycol (PEG). We add 25 mL of 40% PEG solution...
  7. Screening for Successful Genome Editing with Digital PCR

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    Blog Post
    ...resulting from non-homologous end joining (NHEJ) (2-5), depending on your desired genome edit. Since exploitation...each sample can be quickly calculated (details in [2]) and can be used to quantify rates of mutation in... have been reported for mismatch nuclease assays (2, 6, 7). Ability to distinguish between mono-allelic...assays, in addition to testing their performance (2). Remember that although digital PCR is a great screening...PMID: 24509632. PubMed Central PMCID: PMC4063274. 2. Findlay, S. D., Vincent, K. M., Berman, J. R. & Postovit...mono-allelic and bi-allelic mutations: Perhaps the most noteworthy advantage of digital PCR assays is that they ...
  8. DAP arrays for multiplex gene editing

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    Blog Post
    ... as muscular dystrophies, heart disease, and type 2 diabetes, across four different types of human cells...(#186715) and hCtRNA_VT (#186716).   Fig. 2: Comparison of previous representative dual-gRNA architectures... in a variety of cells and organisms. However, lengthy genetic payloads and insufficient transcription...assembled tRNA-gRNAs is placed downstream of a lengthy promoter, which is used to drive the expression...efficient MBE with such a strategy, the need for lengthy promoters, such as U6 or EF1α promoters, has limited...previous tRNA-gRNA arrays, that rely on an upstream lengthy promoter to express the gRNA array, DAP arrays ...
  9. Deep Mutational Scanning with One Pot Saturation Mutagenesis

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    Blog Post
    ... that results in a polymerase error rate of up to 2% per nucleotide position. The “sloppy” PCR reaction...modifications: 1) increased concentration of Taq polymerase, 2) increased PCR extension time, 3) increased concentration...to synthesize a complementary mutant strand (step 2 and 3). Then the uracil-labeled template strand is...restrict site matter? Most of the time no, but there are 2 instances where it does. 1) When the backbone vector...ExoIII. See the below figure for an example like this. 2) When your gene of interest has a BbvCI site(s). If...saturation mutagenesis method are 1) ease of use, 2) time savings, and 3) ability to perform single site...PMID: 23766324. PubMed Central PMCID: PMC4409680. 2. Mccullum, E. O., Williams, B. A., Zhang, J., & Chaput...
  10. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...fluorescent state make them useful for 2-color imaging with green PA-FPs. Table 2: Properties of Selected Reversible... conventional FP imaging. PA-FPs are divided into 2 main categories - those with irreversible and reversible... forms of the chromophore which contribute to the 2 peaks of wtGFP’s excitation spectrum- a major 397 ... / N99K / S148L / I165V / Q167P / L169V / I203R) (2) and PA-mRFP1(S146H / I161V / I197H) (3) were derived...with two distinct wavelengths of light (see Table 2). The best known reversible PA-FP is the naturally... above background. The majority of PA-FPs come in 2 flavours- monomeric and tetrameric. Tetrameric PA-...most PA-FPs, but, in 2015, the Looger Lab designed 2 variants of EosFP that can withstand these conditions...
Showing: 31 - 40 of 136 results