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We narrowed to 432 results for: sequence

Showing: 121 - 130 of 432 results
  1. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...such as this "CRISPR 101" post. Next-generation sequencing technologies are the gold standard but they remain...assays can be designed to detect either a donor sequence integrated through homology-directed repair (HDR...designed to perfectly complement the wild type sequence and is not capable of binding a mutated target... screening tool, it still requires target DNA sequencing to confirm the exact nature of your mutations...great description of the plethora of available sequencing options can be found here. Good luck with your...
  2. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...knock-in long sequences with reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally...M-CREATE) dCas9-SSAP with reduced error rate for long sequence knock-ins Visual barcodes for live cell clonal...  dCas9-SSAP with reduced error rate for long sequence knock-ins by: Lucie Wilson Le Cong’s lab has developed...SSAP will make the editing of kilobase-scaled sequences more efficient and less risky.   Fig. 3...
  3. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...clonal dynamics over time using next generation sequencing (NGS). The library was developed when Carrie ...population of cells with specifc, easily read sequences that can be used to measure their clonal abundance...barcoded clone can then be monitored over time by sequencing the barcodes in the population (all barcodes ...primers). The abundance of a particular barcode sequence then corresponds to the abundance of a specific...clone; lineage tracing is performed via barcode sequencing. Using the ClonTracer library, Bhang et al. found...
  4. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    Published
    Jan. 12, 2017, 3:30 p.m.
    ... a rapid and highly efficient way to move DNA sequences into multiple vector systems. With the appropriate...scientists can easily clone in their desired DNA sequences. Gateway technology relies on the two reactions...add attB sites to either end of the KRAS coding sequence. If you choose this strategy, it’s important to...protein expression elements (ribosome recognition sequences, start codon, stop codons, reading frame considerations...verify the integrity of your expression clone via sequencing or restriction digest! Then, you can transform...Once you generate the entry clone with your DNA sequence of interest, you can move this DNA fragment across...
  5. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ...mariner), as a method of tagmentation for next-gen sequencing pipelines (e.g. using Tn5), and as knock-in tools...A CRISPR RNA (crRNA) comprising a 32 nt spacer sequence specifying the target site. Four proteins (TniQ...interest flanked by ~50-150 bp transposon end sequences (R and L ends).  Figure 1: (A) Components.... Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although 5’-CC-3’...complexes survey the genome and bind target DNA sequences using RNA-DNA base pairing. Separately, TnsABC...overnight Screen colonies phenotypically or by PCR/sequencing While CRISPR-transposons are powerful and simple...
  6. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ... cells. Cas13 enzymes also don’t require a PAM sequence at the target locus, making them more flexible...given single base protospacer flanking site (PFS) sequence, but orthologs like LwaCas13a do not require a... target RNA from bases 12-26 of a 30 nt target sequence. For the editing portion of the protein, they ...designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found that 50...target effects observed in whole-transcriptome RNA-sequencing, the authors performed rational mutagenesis and...RESCUE further expands the RNA editing toolkit and sequences that can be edited.   As discussed above, the ...
  7. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    Published
    Oct. 27, 2015, 3 p.m.
    ...positioning of the FP sequence relative to cell compartment targeting sequences, which often have absolute...requirements. For example, a KDEL ER retrieval sequence only functions at the extreme C-terminus [3]. ...Additionally, FPs that encode N-glycosylation consensus sequences are modified with the addition of N-glycans [9...using FPs containing two cysteine residues and a sequence intended to localize them to the secretory pathway...cysteine residues and N-glycosylation consensus sequences in popular FPs. This generated a palette of inert...others (5, 6, 7, 10, 11) highlight the very real consequences of misfolded dark FP fusions including erroneous...
  8. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ...this gap, the Rat Genome Sequencing Project conducted a large scale sequencing and concluded that nearly... fully sequenced by the year 2000, becoming the first plant to have its entire genome sequenced (Koornneff... limited at first, advances in axolotl genome sequencing have vastly expanded axolotl research. One of...of the most significant advances is the sequencing of the entire axolotl genome, revealing an abundance..., R., Weinstock, G., Metzker, M. et al. Genome sequence of the Brown Norway rat yields insights into mammalian...
  9. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    Published
    Nov. 2, 2021, 1:15 p.m.
    ...the same sequence to flank a transgene (homologous pairs) or used different scrambled sequences to flank...Recombinase-dependent vectors utilize pairs of specific sequences, lox (recognized by Cre) or frt (recognized by...used a lox-STOP-lox (LSL) or frt-STOP-frt (FSF) sequence prior to an ATG start codon to prevent transcription...recognized by Cre, but they are comprised of different sequences (loxp and lox2272) and can not recombine with ...phenomena by analyzing the plasmid and viral DNA sequences and expression profile at multiple steps from ...containing a DIO/FLEx cassette using next-generation sequencing (NGS) (Guerin, K. 2020). In general, we observe...Mitsopoulos A, Fan M (2020) A Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity ...
  10. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ...initiation sequence known as the Species-Independent Translational Sequence (SITS). This sequence bypasses... via DNA sequencing or other readout. Cellular memory is accomplished by the use of sequence-specific ...in series with unique spacer sequences (pMemoryArray) gives a DNA sequence capable of recording 11 bits...shuttled out of the nucleus due to the nuclear export sequence. Treatment with rapamycin induces Cas9(N)-FRB-... the presence of multiple copies of targeting sequences. Scientists have adopted and employed this concept...inducible expression, loxP sites, H2B, P2A or PEST2D sequences. Several pre-constructed promoter-selection marker...generating plasmids with varying numbers of repeated sequences or inserts, which can be advantageous for imaging...
Showing: 121 - 130 of 432 results