nebuffer 2  (New England Biolabs)

Journal: Molecular Ecology Resources

Article Title: Development and Laboratory Validation of a Field‐Deployable CRISPR ‐Cas12a eDNA Assay for Phylogeographic Lineage Detection in Arctic Char ( Salvelinus alpinus )

doi: 10.1111/1755-0998.70125

Figure Lengend Snippet: Influence of buffer concentrations on PAM site tolerance by LbCas12a. Titration of NEBuffer 2.1 indicated that a 5× concentration most effectively differentiated between lineages by modulating LbCas12a's tolerance for suboptimal PAM sites in Atlantic Subclade lineages 2–4, emphasizing the importance of buffer conditions in assay specificity. A total of 1 ng of tissue extracted DNA was used in each RPA‐CRISPR‐Cas reaction.

Article Snippet: The effect of NEBuffer 2.1 concentration on LbCas12a assay performance was systematically evaluated using buffer concentrations ranging from 1× to 6× (Figure ).

Techniques: Titration, Concentration Assay, CRISPR

Journal: Molecular Ecology Resources

Article Title: Development and Laboratory Validation of a Field‐Deployable CRISPR ‐Cas12a eDNA Assay for Phylogeographic Lineage Detection in Arctic Char ( Salvelinus alpinus )

doi: 10.1111/1755-0998.70125

Figure Lengend Snippet: Influence of buffer concentrations on PAM site tolerance by LbCas12a. Titration of NEBuffer 2.1 indicated that a 5× concentration most effectively differentiated between lineages by modulating LbCas12a's tolerance for suboptimal PAM sites in Atlantic Subclade lineages 2–4, emphasizing the importance of buffer conditions in assay specificity. A total of 1 ng of tissue extracted DNA was used in each RPA‐CRISPR‐Cas reaction.

Article Snippet: Buffer optimisation was similarly assessed by varying NEBuffer 2.1 concentrations, with the optimal condition defined as the one yielding the highest specificity without loss of sensitivity.

Techniques: Titration, Concentration Assay, CRISPR