Sequel binding and internal control kit 2

Manufactured by Pacific Biosciences

The Sequel Binding and Internal Control Kit 2.1 is a laboratory equipment product from Pacific Biosciences. It is designed to facilitate the binding of DNA samples to the SMRT Cell used in the Sequel System for long-read sequencing. The kit includes reagents and consumables required for this process.

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Lab products found in correlation

Smrt link v5, by Pacific Biosciences (3 mentions) Smrtbell template prep kit 1.0 spv3, by Pacific Biosciences (3 mentions) Sequel sequencing kit 2, by Pacific Biosciences (3 mentions) Tapestation instrument, by Agilent Technologies (2 mentions) Ampure pb bead, by Pacific Biosciences (1 mentions)

Spelling variants of this product

Sequel II (38 citations) Sequel II Binding Kit 2.0 (22 citations) Sequel Binding Kit 2.0 (8 citations)

3 protocols using sequel binding and internal control kit 2

SMRT Sequencing of Captured Libraries

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The pretarget captured sequencing libraries with Illumina format adapters were pooled and subjected to single‐molecule real‐time (SMRT) sequencing template construction using a SMRTbell Template Prep Kit 1.0—SPv3 (Pacific Biosciences). The posttarget captured Illumina sequencing libraries were similarly pooled and further subjected to SMRT sequencing library construction. The amplicon template preparation and sequencing protocol was used, with minor modifications: DNA was purified with 1.8× AMPure PB beads, and library size was estimated using a TapeStation instrument (Agilent). Sequencing primer annealing and polymerase binding conditions were calculated with the SMRT Link v5.1.0 software (Pacific Biosciences). Briefly, sequencing primer v3 was annealed to the sequencing template, and then polymerase was bound to templates using a Sequel Binding and Internal Control Kit 2.1 (Pacific Biosciences). Sequencing was performed on a Sequel SMRT Cell 1M v2. Sequencing movies were collected on the Sequel system for 10 hours with a Sequel Sequencing Kit 2.1 (Pacific Biosciences).

Vong J.S., Jiang P., Cheng S., Lee W., Tsang J.C., Leung T., Chan K.C., Chiu R.W, & Lo Y.M. (2019). Enrichment of fetal and maternal long cell‐free DNA fragments from maternal plasma following DNA repair. Prenatal Diagnosis, 39(2), 88-99.

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PacBio Single-Molecule Sequencing

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Sequencing templates were constructed using SMRTbell Template Prep Kit 1.0 - SPv3 (Pacific Biosciences) according to the manufacturer’s instructions, except that the amplicon templates were purified with AMPure PB beads (Pacific Biosciences). Sequencing primer annealing and polymerase binding conditions were calculated with SMRT Link v5.1.0 software (Pacific Biosciences). Briefly, sequencing primer v3 was annealed to sequencing template, then polymerase was bound to templates using Sequel Binding and Internal Control Kit 2.1 (Pacific Biosciences). Sequencing was performed on a Sequel SMRT Cell 1M v2 for each template. Each sequencing movie was collected on the Sequel system for 10 h with Sequel sequencing kit 2.1 (Pacific Biosciences).

Serpas L., Chan R.W., Jiang P., Ni M., Sun K., Rashidfarrokhi A., Soni C., Sisirak V., Lee W.S., Cheng S.H., Peng W., Chan K.C., Chiu R.W., Reizis B, & Lo Y.M. (2018). Dnase1l3 deletion causes aberrations in length and end-motif frequencies in plasma DNA. Proceedings of the National Academy of Sciences of the United States of America, 116(2), 641-649.

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Highly Accurate PacBio Sequencing Protocol

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Sequencing templates were constructed using SMRTbell Template Prep Kit 1.0-SPv3 (Pacific Biosciences). The amplicon template preparation and sequencing protocol was used, with minor modifications: DNA was purified with 1.8× AMPure PB beads, and library size was estimated using TapeStation instrument (Agilent). Sequencing primer annealing and polymerase binding conditions were calculated with SMRT Link v.5.1.0 software (Pacific Biosciences). Briefly, sequencing primer v.3 was annealed to sequencing template, then the polymerase was bound to templates using Sequel Binding and Internal Control Kit 2.1 (Pacific Biosciences). Sequencing was performed on a Sequel SMRT Cell 1M v.2 for each template. Sequencing movie was collected on the Sequel system for 10 h with Sequel sequencing kit 2.1 (Pacific Biosciences).

Chan R.W., Serpas L., Ni M., Volpi S., Hiraki L.T., Tam L.S., Rashidfarrokhi A., Wong P.C., Tam L.H., Wang Y., Jiang P., Cheng A.S., Peng W., Han D.S., Tse P.P., Lau P.K., Lee W.S., Magnasco A., Buti E., Sisirak V., AlMutairi N., Chan K.C., Chiu R.W., Reizis B, & Lo Y.M. (2020). Plasma DNA Profile Associated with DNASE1L3 Gene Mutations: Clinical Observations, Relationships to Nuclease Substrate Preference, and In Vivo Correction. American Journal of Human Genetics, 107(5), 882-894.

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