Journal: Nature protocols

Article Title: RNase H-dependent PCR-enabled T Cell Receptor sequencing (rhTCRseq) for Highly Specific and Efficient Targeted Sequencing of T Cell Receptor mRNA for Single-Cell and Repertoire Analysis

doi: 10.1038/s41596-019-0195-x

Figure Lengend Snippet:

Article Snippet: Prepare the following reverse transcriptase (RT) mix and keep on ice: Component Volume for 96-well plate with overage (μL) Volume for 384-well plate with overage (μL) Final concentration in RT reaction NEBNext Single Cell RT Buffer (4×) 120 207.5 1× AAPV Proteinase K inhibitor (25 mM) 9.6 16.6 0.5 mM NEBNext Template Switching Oligo 24 41.5 - NEBNext Single Cell RT Enzyme Mix 48 83 - Water 38.4 66.4 - Total volume 240 415 Open in a separate window With the plate in cooling block on the deck of the Mantis, use Mantis to dispense 1 μL of RT mix to each of the wells if using a 384-well plate, or 2 μL per well for a 96-well plate.

Techniques: Concentration Assay, Lysis

Journal: Nature protocols

Article Title: RNase H-dependent PCR-enabled T Cell Receptor sequencing (rhTCRseq) for Highly Specific and Efficient Targeted Sequencing of T Cell Receptor mRNA for Single-Cell and Repertoire Analysis

doi: 10.1038/s41596-019-0195-x

Figure Lengend Snippet:

Article Snippet: Prepare the following reverse transcriptase (RT) mix and keep on ice: Component Volume for 96-well plate with overage (μL) Volume for 384-well plate with overage (μL) Final concentration in RT reaction NEBNext Single Cell RT Buffer (4×) 120 207.5 1× AAPV Proteinase K inhibitor (25 mM) 9.6 16.6 0.5 mM NEBNext Template Switching Oligo 24 41.5 - NEBNext Single Cell RT Enzyme Mix 48 83 - Water 38.4 66.4 - Total volume 240 415 Open in a separate window With the plate in cooling block on the deck of the Mantis, use Mantis to dispense 1 μL of RT mix to each of the wells if using a 384-well plate, or 2 μL per well for a 96-well plate.

Techniques: Concentration Assay