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Illumina Inc barcoded reverse primer
Barcoded Reverse Primer, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 82/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/barcoded reverse primer/product/Illumina Inc
Average 82 stars, based on 1 article reviews
Price from $9.99 to $1999.99
barcoded reverse primer - by Bioz Stars, 2019-10
82/100 stars

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Polymerase Chain Reaction:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Incubation was followed by exonuclease treatment: 0.5μl exonuclease I (New England Biolabs), 0.5μl exonuclease III (New England Biolabs), 0.2μl 10x ampligase buffer (Illumina), and 0.8μl H2 O was added to the (cooled) capture samples (consecutively incubated at 37°C and 95°C for 45 and 2 min, respectively). .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ). .. Sequencing of the barcoded samples was performed using the Illumina NextSeq500 system, with 2 × 151‐bp paired‐end reads; smMIP libraries required spike‐in of custom primers as described previously [O'Roak et al., ].

Size-exclusion Chromatography:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Incubation was followed by exonuclease treatment: 0.5μl exonuclease I (New England Biolabs), 0.5μl exonuclease III (New England Biolabs), 0.2μl 10x ampligase buffer (Illumina), and 0.8μl H2 O was added to the (cooled) capture samples (consecutively incubated at 37°C and 95°C for 45 and 2 min, respectively). .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ). .. Sequencing of the barcoded samples was performed using the Illumina NextSeq500 system, with 2 × 151‐bp paired‐end reads; smMIP libraries required spike‐in of custom primers as described previously [O'Roak et al., ].

Sequencing:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Paragraph title: Targeted Sequencing of BRCA1 and BRCA2 by smMIPs ... Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ).

Incubation:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Incubation was followed by exonuclease treatment: 0.5μl exonuclease I (New England Biolabs), 0.5μl exonuclease III (New England Biolabs), 0.2μl 10x ampligase buffer (Illumina), and 0.8μl H2 O was added to the (cooled) capture samples (consecutively incubated at 37°C and 95°C for 45 and 2 min, respectively). .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ).

Formalin-fixed Paraffin-Embedded:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Targeted sequencing of BRCA1 and BRCA2 using DNA derived from FFPE OCs was performed as previously described [Weren et al., ], using a slightly modified smMIP capture protocol [O'Roak et al., ; Hiatt et al., ]. .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ).

Modification:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Targeted sequencing of BRCA1 and BRCA2 using DNA derived from FFPE OCs was performed as previously described [Weren et al., ], using a slightly modified smMIP capture protocol [O'Roak et al., ; Hiatt et al., ]. .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ).

Derivative Assay:

Article Title: Novel BRCA1 and BRCA2 Tumor Test as Basis for Treatment Decisions and Referral for Genetic Counselling of Patients with Ovarian Carcinomas
Article Snippet: Targeted sequencing of BRCA1 and BRCA2 using DNA derived from FFPE OCs was performed as previously described [Weren et al., ], using a slightly modified smMIP capture protocol [O'Roak et al., ; Hiatt et al., ]. .. Subsequently, 10μl of the sample was used to perform a PCR reaction by adding 1.3μl of barcoded reverse primer (10μM; Illumina), 12.5μl 2x iProof (BioRad Laboratories, Veenendaal, the Netherlands), 0.125μl forward primer (100μM; Illumina), and 1.075μl H2 O (final volume: 25μl; PCR program: 98°C, 30 sec – 24x [98°C, 10 sec ‐ 60°C, 30 sec ‐ 72°C, 30 sec] ‐ 72°C, 2 min ‐ 4°C, ∞ ).

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  • 87
    Illumina Inc barcoded reverse primers
    Generating natively paired libraries for screening and next-generation sequencing (NGS). a V H and V L domains from each encapsulated B cell are amplified with specific primer sets and paired in-frame via complementary overhangs (green). A nested PCR with V H FR1 and V L FR4 primers generates full-length scFv with overhangs (purple) to enable <t>barcoded</t> paired MiSeq sequencing. V H and V L FR3, CDR3 and FR4 are sequenced with R2 and R1 primers, respectively, while the standard P5-specific primer provides the index read to enable demultiplexing. The amplicons can be cleaved of adapter sequences via Sfi1/Not1 restriction sites for subcloning into expression or phagemid vectors for library generation. b , c NGS data from a representative dataset where scFv libraries from one million stimulated B cells were generated in either emulsion or combinatorial fashion. b Emulsion libraries favored a 1:1 V L :V H ratio whereas combinatorial libraries were scrambled. c In the case where multiple pairings were seen, the dominant partner accounted for 96% of sequences in the emulsion library, whereas partners were more evenly distributed in the combinatorial library
    Barcoded Reverse Primers, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/barcoded reverse primers/product/Illumina Inc
    Average 87 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    barcoded reverse primers - by Bioz Stars, 2019-10
    87/100 stars
      Buy from Supplier

    86
    Illumina Inc barcoded its2 reverse primers
    Generating natively paired libraries for screening and next-generation sequencing (NGS). a V H and V L domains from each encapsulated B cell are amplified with specific primer sets and paired in-frame via complementary overhangs (green). A nested PCR with V H FR1 and V L FR4 primers generates full-length scFv with overhangs (purple) to enable <t>barcoded</t> paired MiSeq sequencing. V H and V L FR3, CDR3 and FR4 are sequenced with R2 and R1 primers, respectively, while the standard P5-specific primer provides the index read to enable demultiplexing. The amplicons can be cleaved of adapter sequences via Sfi1/Not1 restriction sites for subcloning into expression or phagemid vectors for library generation. b , c NGS data from a representative dataset where scFv libraries from one million stimulated B cells were generated in either emulsion or combinatorial fashion. b Emulsion libraries favored a 1:1 V L :V H ratio whereas combinatorial libraries were scrambled. c In the case where multiple pairings were seen, the dominant partner accounted for 96% of sequences in the emulsion library, whereas partners were more evenly distributed in the combinatorial library
    Barcoded Its2 Reverse Primers, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 86/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/barcoded its2 reverse primers/product/Illumina Inc
    Average 86 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    barcoded its2 reverse primers - by Bioz Stars, 2019-10
    86/100 stars
      Buy from Supplier

    93
    Illumina Inc barcoded illumina primers
    Generating natively paired libraries for screening and next-generation sequencing (NGS). a V H and V L domains from each encapsulated B cell are amplified with specific primer sets and paired in-frame via complementary overhangs (green). A nested PCR with V H FR1 and V L FR4 primers generates full-length scFv with overhangs (purple) to enable <t>barcoded</t> paired MiSeq sequencing. V H and V L FR3, CDR3 and FR4 are sequenced with R2 and R1 primers, respectively, while the standard P5-specific primer provides the index read to enable demultiplexing. The amplicons can be cleaved of adapter sequences via Sfi1/Not1 restriction sites for subcloning into expression or phagemid vectors for library generation. b , c NGS data from a representative dataset where scFv libraries from one million stimulated B cells were generated in either emulsion or combinatorial fashion. b Emulsion libraries favored a 1:1 V L :V H ratio whereas combinatorial libraries were scrambled. c In the case where multiple pairings were seen, the dominant partner accounted for 96% of sequences in the emulsion library, whereas partners were more evenly distributed in the combinatorial library
    Barcoded Illumina Primers, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/barcoded illumina primers/product/Illumina Inc
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    barcoded illumina primers - by Bioz Stars, 2019-10
    93/100 stars
      Buy from Supplier

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    Generating natively paired libraries for screening and next-generation sequencing (NGS). a V H and V L domains from each encapsulated B cell are amplified with specific primer sets and paired in-frame via complementary overhangs (green). A nested PCR with V H FR1 and V L FR4 primers generates full-length scFv with overhangs (purple) to enable barcoded paired MiSeq sequencing. V H and V L FR3, CDR3 and FR4 are sequenced with R2 and R1 primers, respectively, while the standard P5-specific primer provides the index read to enable demultiplexing. The amplicons can be cleaved of adapter sequences via Sfi1/Not1 restriction sites for subcloning into expression or phagemid vectors for library generation. b , c NGS data from a representative dataset where scFv libraries from one million stimulated B cells were generated in either emulsion or combinatorial fashion. b Emulsion libraries favored a 1:1 V L :V H ratio whereas combinatorial libraries were scrambled. c In the case where multiple pairings were seen, the dominant partner accounted for 96% of sequences in the emulsion library, whereas partners were more evenly distributed in the combinatorial library

    Journal: Communications Biology

    Article Title: Recombinant human B cell repertoires enable screening for rare, specific, and natively paired antibodies

    doi: 10.1038/s42003-017-0006-2

    Figure Lengend Snippet: Generating natively paired libraries for screening and next-generation sequencing (NGS). a V H and V L domains from each encapsulated B cell are amplified with specific primer sets and paired in-frame via complementary overhangs (green). A nested PCR with V H FR1 and V L FR4 primers generates full-length scFv with overhangs (purple) to enable barcoded paired MiSeq sequencing. V H and V L FR3, CDR3 and FR4 are sequenced with R2 and R1 primers, respectively, while the standard P5-specific primer provides the index read to enable demultiplexing. The amplicons can be cleaved of adapter sequences via Sfi1/Not1 restriction sites for subcloning into expression or phagemid vectors for library generation. b , c NGS data from a representative dataset where scFv libraries from one million stimulated B cells were generated in either emulsion or combinatorial fashion. b Emulsion libraries favored a 1:1 V L :V H ratio whereas combinatorial libraries were scrambled. c In the case where multiple pairings were seen, the dominant partner accounted for 96% of sequences in the emulsion library, whereas partners were more evenly distributed in the combinatorial library

    Article Snippet: A final scale-up PCR was performed using common forward (Illu_scaleup_F) and barcoded reverse primers (Illu_R_N50X) to enable library construction and Illumina sequencing (Supplementary Table ).

    Techniques: Next-Generation Sequencing, Amplification, Nested PCR, Sequencing, Subcloning, Expressing, Generated