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t1130l  (New England Biolabs)


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    New England Biolabs t1130l
    T1130l, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 286 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t1130l/product/New England Biolabs
    Average 99 stars, based on 286 article reviews
    t1130l - by Bioz Stars, 2026-02
    99/100 stars

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    ssDNA isolation and library preparation (A) Prior to the addition of fuel, the peptide, polyU RNA and ssDNA molecules are mixed in solution, and do not form supramolecular assemblies. When fuel (EDC) is added, phase separation occurs, forming coacervate droplets that sequester the ssDNA molecules from the polymer-poor phase. The phases are separated via centrifugation to yield the polymer-poor and polymer-rich phases, from which the ssDNA populations are isolated. For a complete characterization of this droplet system, please refer to Donau et al., 2020. (B) Library preparation relies on the orientation-specific ligation of two adapter molecules to the recovered ssDNA oligonucleotides. After the ligation, the sequencing primers are added in a PCR using primers with overhangs. (C) Urea-PAGE is performed following library preparation. R = Reference, S = Supernatant, D = Droplets. ‘Input’ corresponds to the samples obtained from the droplets, while the ‘R’ sample is the naïve input library. ‘Ligation’ corresponds to the product of the first step in B and shows a clear band (red star) corresponding to the expected full-length ligation product, as well as a weaker smear (green stars) corresponding to the two partially adapter-ligated products. Bands at 45 nt correspond to the adapters and splint, and bands at ∼30 correspond to unreacted input ssDNA. ‘Product’ shows the full-length sequencing library, with the correct PCR band being marked with a red star. Several additional bands are visible, including two prominent bands (blue stars), which are located approximately 30 nt above and below the main PCR band. These correspond to the no-insert (adapter dimer) and double-insert ligation products. The smear and bands below correspond to indexing primers. (E) Electropherogram of a TapeStation run of a prepared library. Only the peak corresponding to the correct library size (∼160 bp) is used to calculate the correct library dilution for loading onto the sequencer.

    Journal: STAR Protocols

    Article Title: Protocol for the recovery and deep sequencing of short ssDNA pools from transient, fuel-dependent coacervate droplets

    doi: 10.1016/j.xpro.2025.104293

    Figure Lengend Snippet: ssDNA isolation and library preparation (A) Prior to the addition of fuel, the peptide, polyU RNA and ssDNA molecules are mixed in solution, and do not form supramolecular assemblies. When fuel (EDC) is added, phase separation occurs, forming coacervate droplets that sequester the ssDNA molecules from the polymer-poor phase. The phases are separated via centrifugation to yield the polymer-poor and polymer-rich phases, from which the ssDNA populations are isolated. For a complete characterization of this droplet system, please refer to Donau et al., 2020. (B) Library preparation relies on the orientation-specific ligation of two adapter molecules to the recovered ssDNA oligonucleotides. After the ligation, the sequencing primers are added in a PCR using primers with overhangs. (C) Urea-PAGE is performed following library preparation. R = Reference, S = Supernatant, D = Droplets. ‘Input’ corresponds to the samples obtained from the droplets, while the ‘R’ sample is the naïve input library. ‘Ligation’ corresponds to the product of the first step in B and shows a clear band (red star) corresponding to the expected full-length ligation product, as well as a weaker smear (green stars) corresponding to the two partially adapter-ligated products. Bands at 45 nt correspond to the adapters and splint, and bands at ∼30 correspond to unreacted input ssDNA. ‘Product’ shows the full-length sequencing library, with the correct PCR band being marked with a red star. Several additional bands are visible, including two prominent bands (blue stars), which are located approximately 30 nt above and below the main PCR band. These correspond to the no-insert (adapter dimer) and double-insert ligation products. The smear and bands below correspond to indexing primers. (E) Electropherogram of a TapeStation run of a prepared library. Only the peak corresponding to the correct library size (∼160 bp) is used to calculate the correct library dilution for loading onto the sequencer.

    Article Snippet: Monarch Spin PCR & DNA Cleanup Kit , New England Biolabs , T1130L.

    Techniques: Isolation, Polymer, Centrifugation, Ligation, Sequencing