repli g single cell kit  (Qiagen)


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    Name:
    REPLI g Single Cell Kit
    Description:
    For highly uniform whole genome amplification WGA from single cells or limited sample material Kit contents Qiagen REPLI g Single Cell Kit 24 WGA rxns 2 to 1000 Cells Input Amount 15 min Hands on Time 8 to 16 hr Reaction Time 40g rxns Yields MDA Technology Tube Format For Highly Uniform Whole Genome Amplification WGA from Single Cells or Limited Sample Includes REPLI g sc Polymerase Buffers and Reagents Benefits WGA from single cell material with complete genome coverage Unbiased amplification of genomic loci due to MDA technology Optimized for use with new technologies including NGS Consistent yields of up to 40 µg average product length 10 kb Novel tool for cancer research stem cell research or metagenomics
    Catalog Number:
    150343
    Price:
    529
    Category:
    REPLI g Single Cell Kit
    Buy from Supplier


    Structured Review

    Qiagen repli g single cell kit
    REPLI g Single Cell Kit
    For highly uniform whole genome amplification WGA from single cells or limited sample material Kit contents Qiagen REPLI g Single Cell Kit 24 WGA rxns 2 to 1000 Cells Input Amount 15 min Hands on Time 8 to 16 hr Reaction Time 40g rxns Yields MDA Technology Tube Format For Highly Uniform Whole Genome Amplification WGA from Single Cells or Limited Sample Includes REPLI g sc Polymerase Buffers and Reagents Benefits WGA from single cell material with complete genome coverage Unbiased amplification of genomic loci due to MDA technology Optimized for use with new technologies including NGS Consistent yields of up to 40 µg average product length 10 kb Novel tool for cancer research stem cell research or metagenomics
    https://www.bioz.com/result/repli g single cell kit/product/Qiagen
    Average 90 stars, based on 73 article reviews
    Price from $9.99 to $1999.99
    repli g single cell kit - by Bioz Stars, 2020-01
    90/100 stars

    Images

    1) Product Images from "Comparative study of whole genome amplification and next generation sequencing performance of single cancer cells"

    Article Title: Comparative study of whole genome amplification and next generation sequencing performance of single cancer cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.10701

    Plots of CNA profiles along the whole genome (x axis) ( A ) CNA profile of unamplified DNA from unfixed cells. ( B – G ) plots of CNAs in single SK-BR-3 cells, obtained from EDTA-preserved blood. ( H , I ) CNA profiles of individual CTCs, obtained from EDTA-preserved blood of the same breast cancer patient. WGA kits: (B, E) Ampli1; (C, F, H, I) PicoPlex; (D, G) REPLI-g.
    Figure Legend Snippet: Plots of CNA profiles along the whole genome (x axis) ( A ) CNA profile of unamplified DNA from unfixed cells. ( B – G ) plots of CNAs in single SK-BR-3 cells, obtained from EDTA-preserved blood. ( H , I ) CNA profiles of individual CTCs, obtained from EDTA-preserved blood of the same breast cancer patient. WGA kits: (B, E) Ampli1; (C, F, H, I) PicoPlex; (D, G) REPLI-g.

    Techniques Used: Whole Genome Amplification

    Distribution of identified known SNPs between datasets ( A ) Known SNPs identified in single cells, amplified with Ampli1, PicoPlex, and REPLI-g WGA kits and obtained from EDTA-preserved blood in comparison to unamplified DNA. ( B ) Known SNPs identified in single cells, amplified with Ampli1 or PicoPlex and obtained from EDTA- and CellSave-preserved blood in comparison to unamplified DNA from unfixed cells. ( C ) Known SNPs identified in single CTCs, amplified with PicoPlex in comparison to each other.
    Figure Legend Snippet: Distribution of identified known SNPs between datasets ( A ) Known SNPs identified in single cells, amplified with Ampli1, PicoPlex, and REPLI-g WGA kits and obtained from EDTA-preserved blood in comparison to unamplified DNA. ( B ) Known SNPs identified in single cells, amplified with Ampli1 or PicoPlex and obtained from EDTA- and CellSave-preserved blood in comparison to unamplified DNA from unfixed cells. ( C ) Known SNPs identified in single CTCs, amplified with PicoPlex in comparison to each other.

    Techniques Used: Amplification, Whole Genome Amplification

    2) Product Images from "Comparative study of whole genome amplification and next generation sequencing performance of single cancer cells"

    Article Title: Comparative study of whole genome amplification and next generation sequencing performance of single cancer cells

    Journal: Oncotarget

    doi: 10.18632/oncotarget.10701

    Plots of CNA profiles along the whole genome (x axis) ( A ) CNA profile of unamplified DNA from unfixed cells. ( B – G ) plots of CNAs in single SK-BR-3 cells, obtained from EDTA-preserved blood. ( H , I ) CNA profiles of individual CTCs, obtained from EDTA-preserved blood of the same breast cancer patient. WGA kits: (B, E) Ampli1; (C, F, H, I) PicoPlex; (D, G) REPLI-g.
    Figure Legend Snippet: Plots of CNA profiles along the whole genome (x axis) ( A ) CNA profile of unamplified DNA from unfixed cells. ( B – G ) plots of CNAs in single SK-BR-3 cells, obtained from EDTA-preserved blood. ( H , I ) CNA profiles of individual CTCs, obtained from EDTA-preserved blood of the same breast cancer patient. WGA kits: (B, E) Ampli1; (C, F, H, I) PicoPlex; (D, G) REPLI-g.

    Techniques Used: Whole Genome Amplification

    Distribution of identified known SNPs between datasets ( A ) Known SNPs identified in single cells, amplified with Ampli1, PicoPlex, and REPLI-g WGA kits and obtained from EDTA-preserved blood in comparison to unamplified DNA. ( B ) Known SNPs identified in single cells, amplified with Ampli1 or PicoPlex and obtained from EDTA- and CellSave-preserved blood in comparison to unamplified DNA from unfixed cells. ( C ) Known SNPs identified in single CTCs, amplified with PicoPlex in comparison to each other.
    Figure Legend Snippet: Distribution of identified known SNPs between datasets ( A ) Known SNPs identified in single cells, amplified with Ampli1, PicoPlex, and REPLI-g WGA kits and obtained from EDTA-preserved blood in comparison to unamplified DNA. ( B ) Known SNPs identified in single cells, amplified with Ampli1 or PicoPlex and obtained from EDTA- and CellSave-preserved blood in comparison to unamplified DNA from unfixed cells. ( C ) Known SNPs identified in single CTCs, amplified with PicoPlex in comparison to each other.

    Techniques Used: Amplification, Whole Genome Amplification

    Related Articles

    Clone Assay:

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: Paragraph title: Laser Microdissection, Whole Genome Amplification (WGA) and Cloning-Sequencing of 16S RNA Genes ... We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells.

    Amplification:

    Article Title: A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama
    Article Snippet: Paragraph title: DNA extraction, amplification, phylogenetic analysis ... DNA was extracted from 1-4 Laboulbeniales thalli using a modified REPLI-g Single Cell Kit (Qiagen, Valencia, CA, USA) protocol [ ].

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: .. DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences). .. MDA amplification with GenomiPhiV2 was performed as described previously ( ).

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells. .. Bacterial 16S rRNA genes were then amplified by PCR using the bacterial specific primer 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and with the universal prokaryotic reverse primer 1492R (5′-GGTTACCTTGTTACGACTT-3′).

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: The PCR amplified DNA was then purified using the Qiagen QIAquick PCR Purification Kit. .. For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Article Title: Robust Preimplantation Genetic Testing of Huntington Disease by Combined Triplet-Primed PCR Analysis of the HTT CAG Repeat and Multi-Microsatellite Haplotyping
    Article Snippet: .. Single cell whole genome amplification (WGA) was performed using either the REPLI-g Single Cell Kit (Qiagen, Germany) or the illustra GenomiPhi™ V2 DNA Amplification Kit (GE Healthcare, United Kingdom) according to respective manufacturer’s instructions. ..

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen). .. The amplified DNA was diluted (1: 100) for PCR assay.

    Article Title: Cancer panel analysis of circulating tumor cells in patients with breast cancer
    Article Snippet: .. The cell pellets that were kept at −80°C were amplified using the REPLI-g Single Cell kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol. .. Briefly, the cell pellets were mixed with a denaturation buffer (included in the kit) and incubated at 65°C for 10 min.

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: .. 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    Whole Genome Amplification:

    Article Title: Sequencing of Treponema pallidum subsp. pallidum from isolate UZ1974 using Anti-Treponemal Antibodies Enrichment: First complete whole genome sequence obtained directly from human clinical material
    Article Snippet: .. Whole genome amplification (WGA) WGA was carried out using Multiple Displacement Amplification with phi 29 polymerase (REPLI-g Single Cell Kit, Qiagen, Hilden, Germany). .. The WGA products were purified using QIAEX II beads according to the manufacturer’s recommendations (Qiagen, Hilden, Germany).

    Article Title: Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics
    Article Snippet: .. Droplet fusion and single-cell whole genome amplification Prior to reagent introduction into the device, an MDA mixture (REPLI-g Single Cell Kit, QIAGEN) was prepared containing 1.5 μL of stop buffer, 1.5 μL of H2 O, 14.5 μL of reaction buffer, 1.0 μL of enzyme mix, 2.5 μL of 10% Tween-20 (1% v/v concentration), and 0.5 μL of 20x Evagreen (Biotium, Fremont, CA, USA), for use in a 21.5-μL reaction volume. .. The MDA mixture was mixed gently, but completely, by vortexing and loaded into the microfluidic device.

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: .. Whole genome amplification WGA was performed on the extracted DNA by two different technologies, GenomePlex® Single Cell Whole Genome Amplification Kit (WGA4) from Sigma and REPLI-g Single Cell Kit from Qiagen. .. For GenomePlex WGA4 kit: 10 μl of DNA was fragmented for 4 min at 99 °C after adding 1 μl of 10× Single Cell Lysis and Fragmentation Buffer.

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: Paragraph title: Single blastomere isolation and whole-genome amplification ... DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences).

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: Paragraph title: Laser Microdissection, Whole Genome Amplification (WGA) and Cloning-Sequencing of 16S RNA Genes ... We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells.

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: .. For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used. .. Briefly, 15 µl template DNA was loaded into a 0.2 ml PCR tube.

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method. .. We hoped that the primase-based whole-genome amplification (WGA) method would result in less bias compared with the random hexamers used in Qiagen’s WGA kit ( ).

    Article Title: Microfluidic droplet platform for ultrahigh-throughput single-cell screening of biodiversity
    Article Snippet: .. Whole-genome amplification was performed using the REPLI-g Single Cell Kit (Qiagen). .. Fragment libraries were prepared using the Ion Xpress Plus Fragment Library Kit and the Ion Xpress Barcode Adapters 1–16 Kit (Life Technologies) according to the manufacturer’s instructions.

    Article Title: Robust Preimplantation Genetic Testing of Huntington Disease by Combined Triplet-Primed PCR Analysis of the HTT CAG Repeat and Multi-Microsatellite Haplotyping
    Article Snippet: .. Single cell whole genome amplification (WGA) was performed using either the REPLI-g Single Cell Kit (Qiagen, Germany) or the illustra GenomiPhi™ V2 DNA Amplification Kit (GE Healthcare, United Kingdom) according to respective manufacturer’s instructions. ..

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: .. Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen). .. The amplified DNA was diluted (1: 100) for PCR assay.

    Article Title: Cancer panel analysis of circulating tumor cells in patients with breast cancer
    Article Snippet: Paragraph title: Whole genome amplification ... The cell pellets that were kept at −80°C were amplified using the REPLI-g Single Cell kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol.

    Cytometry:

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: V(D)J recombination and transgene integration assays Triple positive (tdTomato+ /CD45.2+ / (CD19+B220)+ ) SLRB-iHP B cells (polycistronic SLR was used) in 16 wpt combined spleen cells were sorted by flow cytometry (FACSAria llu SORP cell sorter, Becton Dickinson). .. Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen).

    Incubation:

    Article Title: Single gametophyte sequencing reveals that crossover events differ between sexes in maize
    Article Snippet: Following 15–20 min incubation, the embryo sac was dissociated from the nucellus and was placed into a drop of 0.54 M mannitol by micropipette. .. Then, 5–15 antipodal cells were transferred into one PCR tube containing phosphate-buffered saline buffer from the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany), and also can be stored at −80 °C for a long time for further use.

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used. .. Two microgram of denaturation buffer DLB was added to the DNA and the mixture was incubated at room temperature for 3 min. Later, 3 µl of Stop Solution was added to stop the lysis reaction.

    Article Title: Cancer panel analysis of circulating tumor cells in patients with breast cancer
    Article Snippet: The cell pellets that were kept at −80°C were amplified using the REPLI-g Single Cell kit (Qiagen GmbH, Hilden, Germany) according to the manufacturer's protocol. .. Briefly, the cell pellets were mixed with a denaturation buffer (included in the kit) and incubated at 65°C for 10 min.

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    Modification:

    Article Title: A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama
    Article Snippet: .. DNA was extracted from 1-4 Laboulbeniales thalli using a modified REPLI-g Single Cell Kit (Qiagen, Valencia, CA, USA) protocol [ ]. .. Thalli were often manually cut in 2-3 parts (through the perithecium) using a #10 surgical blade on disposable Bard-Parker handle (Aspen Surgical, Caledonia, MI, USA) to ensure successful lysis.

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: We performed the same set of microfluidic lysis and multiple displacement amplification (MDA) steps as described in our previous work, with one modification. .. Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method.

    Hybridization:

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    Flow Cytometry:

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: V(D)J recombination and transgene integration assays Triple positive (tdTomato+ /CD45.2+ / (CD19+B220)+ ) SLRB-iHP B cells (polycistronic SLR was used) in 16 wpt combined spleen cells were sorted by flow cytometry (FACSAria llu SORP cell sorter, Becton Dickinson). .. Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen).

    Transferring:

    Article Title: Dissecting meiotic recombination based on tetrad analysis by single-microspore sequencing in maize
    Article Snippet: Isolation and lysis of single cells in tetrads We used a thin glass pipette system and Programmable Microinjector PM 2,000 (MDI, South Plainfield, USA) to isolate single tetrads and single microspores, and to destroy the cell wall. .. The cells were then aspirated into PCR tubes filled with PBS buffer from the REPLI-g Single Cell Kit (QIAGEN, Hilden, Germany).

    Infection:

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: Single control MEF cells were SLRB infected tdTomato+ MEF (14 dpi, polycistronic SLR was used). .. Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen).

    Polymerase Chain Reaction:

    Article Title: Dissecting meiotic recombination based on tetrad analysis by single-microspore sequencing in maize
    Article Snippet: .. The cells were then aspirated into PCR tubes filled with PBS buffer from the REPLI-g Single Cell Kit (QIAGEN, Hilden, Germany). ..

    Article Title: A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama
    Article Snippet: DNA was extracted from 1-4 Laboulbeniales thalli using a modified REPLI-g Single Cell Kit (Qiagen, Valencia, CA, USA) protocol [ ]. .. PCR reactions consisted of 13.3 μL of RedExtract Taq polymerase (Sigma-Aldrich, St. Louis, MO, USA), 2.5 μL of each 10 μM primer, 5.7 μL of H2 O, and 1.0 μL of template DNA.

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: Briefly, each blastomere was washed three times in wash medium, and it was transferred into a 0.2-mL PCR tube containing either 4 μL of the transport medium (PBS, supplied as part of the REPLI-g Single Cell kit, Qiagen) or 1.5 μL alkaline lysis buffer (50 mM DTT and 200 mM KOH), depending on the downstream MDA method. .. DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences).

    Article Title: Single gametophyte sequencing reveals that crossover events differ between sexes in maize
    Article Snippet: .. Then, 5–15 antipodal cells were transferred into one PCR tube containing phosphate-buffered saline buffer from the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany), and also can be stored at −80 °C for a long time for further use. .. Whole-genome amplification and sequencing Based on the standard protocol of REPLI-g Single Cell Kit (Qiagen, Hilden, Germany), lysis of female gametophytes and amplification of whole genome (using MDA) was performed.

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells. .. Bacterial 16S rRNA genes were then amplified by PCR using the bacterial specific primer 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and with the universal prokaryotic reverse primer 1492R (5′-GGTTACCTTGTTACGACTT-3′).

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: The PCR amplified DNA was then purified using the Qiagen QIAquick PCR Purification Kit. .. For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen). .. The amplified DNA was diluted (1: 100) for PCR assay.

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: .. 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    DNA Extraction:

    Article Title: A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama
    Article Snippet: Paragraph title: DNA extraction, amplification, phylogenetic analysis ... DNA was extracted from 1-4 Laboulbeniales thalli using a modified REPLI-g Single Cell Kit (Qiagen, Valencia, CA, USA) protocol [ ].

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: .. 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    Multiple Displacement Amplification:

    Article Title: Sequencing of Treponema pallidum subsp. pallidum from isolate UZ1974 using Anti-Treponemal Antibodies Enrichment: First complete whole genome sequence obtained directly from human clinical material
    Article Snippet: .. Whole genome amplification (WGA) WGA was carried out using Multiple Displacement Amplification with phi 29 polymerase (REPLI-g Single Cell Kit, Qiagen, Hilden, Germany). .. The WGA products were purified using QIAEX II beads according to the manufacturer’s recommendations (Qiagen, Hilden, Germany).

    Article Title: Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics
    Article Snippet: .. Droplet fusion and single-cell whole genome amplification Prior to reagent introduction into the device, an MDA mixture (REPLI-g Single Cell Kit, QIAGEN) was prepared containing 1.5 μL of stop buffer, 1.5 μL of H2 O, 14.5 μL of reaction buffer, 1.0 μL of enzyme mix, 2.5 μL of 10% Tween-20 (1% v/v concentration), and 0.5 μL of 20x Evagreen (Biotium, Fremont, CA, USA), for use in a 21.5-μL reaction volume. .. The MDA mixture was mixed gently, but completely, by vortexing and loaded into the microfluidic device.

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: .. DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences). .. MDA amplification with GenomiPhiV2 was performed as described previously ( ).

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: .. Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method. .. We hoped that the primase-based whole-genome amplification (WGA) method would result in less bias compared with the random hexamers used in Qiagen’s WGA kit ( ).

    Isolation:

    Article Title: Dissecting meiotic recombination based on tetrad analysis by single-microspore sequencing in maize
    Article Snippet: Paragraph title: Isolation and lysis of single cells in tetrads ... The cells were then aspirated into PCR tubes filled with PBS buffer from the REPLI-g Single Cell Kit (QIAGEN, Hilden, Germany).

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: Paragraph title: Single blastomere isolation and whole-genome amplification ... DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences).

    Article Title: Single gametophyte sequencing reveals that crossover events differ between sexes in maize
    Article Snippet: Paragraph title: Isolation of antipodal cells from single embryo sac ... Then, 5–15 antipodal cells were transferred into one PCR tube containing phosphate-buffered saline buffer from the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany), and also can be stored at −80 °C for a long time for further use.

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: Laser Microdissection, Whole Genome Amplification (WGA) and Cloning-Sequencing of 16S RNA Genes Small individual Pleurocapsales colonies and associated coccoid and filamentous cells were isolated using a Zeiss PALM MicroBeam apparatus installed in a clean room. .. We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells.

    Article Title: Microfluidic droplet platform for ultrahigh-throughput single-cell screening of biodiversity
    Article Snippet: Total DNA was isolated using a QIAamp DNA Investigator Kit (Qiagen). .. Whole-genome amplification was performed using the REPLI-g Single Cell Kit (Qiagen).

    Microscopy:

    Article Title: Dissecting meiotic recombination based on tetrad analysis by single-microspore sequencing in maize
    Article Snippet: During the isolation on microscope slide, the tetrad samples were submerged in isolation buffer (27% D -sorbitol solution). .. The cells were then aspirated into PCR tubes filled with PBS buffer from the REPLI-g Single Cell Kit (QIAGEN, Hilden, Germany).

    Purification:

    Article Title: Sequencing of Treponema pallidum subsp. pallidum from isolate UZ1974 using Anti-Treponemal Antibodies Enrichment: First complete whole genome sequence obtained directly from human clinical material
    Article Snippet: Whole genome amplification (WGA) WGA was carried out using Multiple Displacement Amplification with phi 29 polymerase (REPLI-g Single Cell Kit, Qiagen, Hilden, Germany). .. The WGA products were purified using QIAEX II beads according to the manufacturer’s recommendations (Qiagen, Hilden, Germany).

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences). .. WGA products were purified using SPRI-beads (AMPure) at 0.8× total reaction volume.

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: The PCR amplified DNA was then purified using the Qiagen QIAquick PCR Purification Kit. .. For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Sequencing:

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: Each sample was processed using a previously developed microfluidic-based minimetagenomic sequencing method ( ). .. Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method.

    Article Title: Microfluidic droplet platform for ultrahigh-throughput single-cell screening of biodiversity
    Article Snippet: Paragraph title: Whole-Genome Sequencing and Analysis. ... Whole-genome amplification was performed using the REPLI-g Single Cell Kit (Qiagen).

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: .. 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

    Mouse Assay:

    Article Title: Reprogramming mouse fibroblasts into engraftable myeloerythroid and lymphoid progenitors
    Article Snippet: Control T or B cells were sorted from spleen of C57BL/6 mice. .. Whole-genome amplification (WGA) of single cell was performed using the REPLI-g Single Cell kit (Qiagen).

    Chromatin Immunoprecipitation:

    Article Title: Microfluidic droplet platform for ultrahigh-throughput single-cell screening of biodiversity
    Article Snippet: Whole-genome amplification was performed using the REPLI-g Single Cell Kit (Qiagen). .. Sequencing of libraries was performed using the genetic analyzer Ion Proton, the Ion PI HI-Q Sequencing 200 Kit, and the Ion PI Chip Kit v2 (Life Technologies) according to the manufacturer’s instructions.

    Sample Prep:

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: Paragraph title: Microfluidic-based minimetagenomic sample preparation. ... Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method.

    Laser Capture Microdissection:

    Article Title: Key Role of Alphaproteobacteria and Cyanobacteria in the Formation of Stromatolites of Lake Dziani Dzaha (Mayotte, Western Indian Ocean)
    Article Snippet: Paragraph title: Laser Microdissection, Whole Genome Amplification (WGA) and Cloning-Sequencing of 16S RNA Genes ... We then used the REPLI-g Single Cell Kit (Qiagen, Hilden, Germany) to amplify whole genomic DNA of the microdissected cells.

    Produced:

    Article Title: Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics
    Article Snippet: Droplet fusion and single-cell whole genome amplification Prior to reagent introduction into the device, an MDA mixture (REPLI-g Single Cell Kit, QIAGEN) was prepared containing 1.5 μL of stop buffer, 1.5 μL of H2 O, 14.5 μL of reaction buffer, 1.0 μL of enzyme mix, 2.5 μL of 10% Tween-20 (1% v/v concentration), and 0.5 μL of 20x Evagreen (Biotium, Fremont, CA, USA), for use in a 21.5-μL reaction volume. .. By using tubes and pumps as mentioned above, the droplets were then re-injected into a microfluidic device where they were spaced by FC-40 oil and fused one-to-one with MDA droplets (diameter: 73 μm) produced on-chip with FC-40 oil containing 0.02% (v/v) Pico-Surf1.

    Concentration Assay:

    Article Title: Massively parallel whole genome amplification for single-cell sequencing using droplet microfluidics
    Article Snippet: .. Droplet fusion and single-cell whole genome amplification Prior to reagent introduction into the device, an MDA mixture (REPLI-g Single Cell Kit, QIAGEN) was prepared containing 1.5 μL of stop buffer, 1.5 μL of H2 O, 14.5 μL of reaction buffer, 1.0 μL of enzyme mix, 2.5 μL of 10% Tween-20 (1% v/v concentration), and 0.5 μL of 20x Evagreen (Biotium, Fremont, CA, USA), for use in a 21.5-μL reaction volume. .. The MDA mixture was mixed gently, but completely, by vortexing and loaded into the microfluidic device.

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: Cell concentration was adjusted to ∼2 million cells per milliliter before loading onto a Fluidigm C1 microfluidic integrated fluidic circuit. .. Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method.

    Alkaline Lysis:

    Article Title: Zygotes segregate entire parental genomes in distinct blastomere lineages causing cleavage-stage chimerism and mixoploidy
    Article Snippet: Briefly, each blastomere was washed three times in wash medium, and it was transferred into a 0.2-mL PCR tube containing either 4 μL of the transport medium (PBS, supplied as part of the REPLI-g Single Cell kit, Qiagen) or 1.5 μL alkaline lysis buffer (50 mM DTT and 200 mM KOH), depending on the downstream MDA method. .. DNA from single blastomeres was whole-genome amplified using commercial MDA kits (REPLI-g Single Cell Kit, Qiagen; and GenomiPhiV2, GE Healthcare Biosciences).

    Lysis:

    Article Title: Dissecting meiotic recombination based on tetrad analysis by single-microspore sequencing in maize
    Article Snippet: Paragraph title: Isolation and lysis of single cells in tetrads ... The cells were then aspirated into PCR tubes filled with PBS buffer from the REPLI-g Single Cell Kit (QIAGEN, Hilden, Germany).

    Article Title: A tripartite survey of hyperparasitic fungi associated with ectoparasitic flies on bats (Mammalia: Chiroptera) in a neotropical cloud forest in Panama
    Article Snippet: DNA was extracted from 1-4 Laboulbeniales thalli using a modified REPLI-g Single Cell Kit (Qiagen, Valencia, CA, USA) protocol [ ]. .. Thalli were often manually cut in 2-3 parts (through the perithecium) using a #10 surgical blade on disposable Bard-Parker handle (Aspen Surgical, Caledonia, MI, USA) to ensure successful lysis.

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection
    Article Snippet: Whole genome amplification WGA was performed on the extracted DNA by two different technologies, GenomePlex® Single Cell Whole Genome Amplification Kit (WGA4) from Sigma and REPLI-g Single Cell Kit from Qiagen. .. For GenomePlex WGA4 kit: 10 μl of DNA was fragmented for 4 min at 99 °C after adding 1 μl of 10× Single Cell Lysis and Fragmentation Buffer.

    Article Title: Hydrogenotrophic methanogenesis in archaeal phylum Verstraetearchaeota reveals the shared ancestry of all methanogens
    Article Snippet: We performed the same set of microfluidic lysis and multiple displacement amplification (MDA) steps as described in our previous work, with one modification. .. Instead of using Qiagen’s REPLI-g Single Cell Kit for MDA on-chip, we adapted the SYGNIS TruePrime MDA chemistry to the microfluidic minimetagenomic method.

    Article Title: Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations. Laboulbeniales hyperparasites (Fungi, Ascomycota) of bat flies: Independent origins and host associations
    Article Snippet: 2.3 DNA extraction, PCR amplification, sequencing DNA was extracted from 1–14 Laboulbeniales thalli using the Extract‐N‐Amp Plant PCR Kit (Sigma‐Aldrich, St. Louis, Missouri) (Haelewaters et al., ) or the REPLI‐g Single Cell Kit (Qiagen, Valencia, California) (Haelewaters et al., ). .. Pretreatments employed with the Extract‐N‐Amp method included a prolonged incubation period at 56°C in 20 μl Extraction Solution up to 24‐hr in a Shake ‘N Bake Hybridization Oven (Boekel Scientific model #136400‐2, Feasterville, Pennsylvania) and mechanically crushing fungal material in a FastPrep FP120 Cell Disrupter (Thermo Fisher Scientific, Waltham, Massachusetts) at 5.5 m/s for 20 s. For about two thirds of our extractions, and as a rule for later extractions, thalli were manually cut in 2 or 3 parts (usually through the perithecium) using a #10 surgical blade on disposable Bard‐Parker handle (Aspen Surgical, Caledonia, Michigan) to ensure successful lysis.

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    Qiagen repli g single cell kit
    Whole genome amplification by using GenomePlex (WGA4) and <t>REPLI-g.</t> a Comparison of two whole genome amplification methods: GenomePlex (WGA4) and REPLI-g. b Workflow of GenomePlex (WGA4) and REPLI-g kits. c Whole Genome Amplified DNA products. GenomePlex (WGA4) or REPLI-g WGA was performed on DNA from both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA was checked using agarose gel electrophoresis. d DNA yield comparison. GenomePlex (WGA4) or REPLI-g WGA was performed on both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA amount was measured using the Qubit. (*: standard protocol. **: increased DNA volume protocol)
    Repli G Single Cell Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 90/100, based on 72 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/repli g single cell kit/product/Qiagen
    Average 90 stars, based on 72 article reviews
    Price from $9.99 to $1999.99
    repli g single cell kit - by Bioz Stars, 2020-01
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    For DNA library construction from single cells for Illumina sequencing applications Kit contents REPLI g sc DNA Polymerase buffers and library prep reagents for 48 whole genome amplification reactions andsubsequent
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    Whole genome amplification by using GenomePlex (WGA4) and REPLI-g. a Comparison of two whole genome amplification methods: GenomePlex (WGA4) and REPLI-g. b Workflow of GenomePlex (WGA4) and REPLI-g kits. c Whole Genome Amplified DNA products. GenomePlex (WGA4) or REPLI-g WGA was performed on DNA from both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA was checked using agarose gel electrophoresis. d DNA yield comparison. GenomePlex (WGA4) or REPLI-g WGA was performed on both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA amount was measured using the Qubit. (*: standard protocol. **: increased DNA volume protocol)

    Journal: NPJ Genomic Medicine

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection

    doi: 10.1038/s41525-017-0034-3

    Figure Lengend Snippet: Whole genome amplification by using GenomePlex (WGA4) and REPLI-g. a Comparison of two whole genome amplification methods: GenomePlex (WGA4) and REPLI-g. b Workflow of GenomePlex (WGA4) and REPLI-g kits. c Whole Genome Amplified DNA products. GenomePlex (WGA4) or REPLI-g WGA was performed on DNA from both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA was checked using agarose gel electrophoresis. d DNA yield comparison. GenomePlex (WGA4) or REPLI-g WGA was performed on both fresh and fixed HCT116 cells following the vendor’s manuals. The amplified DNA amount was measured using the Qubit. (*: standard protocol. **: increased DNA volume protocol)

    Article Snippet: For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Techniques: Whole Genome Amplification, Amplification, Agarose Gel Electrophoresis

    Sequencing comparison in WGA amplified and non-amplified DNA samples. a Mutation detection comparison for REPLI-g WGA. REPLI-g amplified or non-amplified DNA from both fresh and fixed HCT116 cells were subjected to the CRC targeted NGS. The blue color highlights the true mutations detected, while the number inside each cell represents the MAF of the mutation. The light red color highlights the false mutations called. b Gene coverage comparison for WGA. The coverage was compared among the following samples: ① Fresh cells; ② Fixed cells; ③ Fresh cells + REPLI-g; ④ Fixed cells + REPLI-g. ⑤ Fixed cells+WGA4. The reads were aligned using BWA-MEM and coverage was computed using GATK’s DepthOfCoverage and in-house scripts. The percentage of bases covered by at least 10 reads with minimum base quality score Q30 (%_bases_above_10) was calculated using data obtained from DepthOfCoverage and in-house scripts and summarized in the table. (*) In the case of GenomePlex (WGA4), the plotted data was trimmed of 10 bp at the beginning and 40 bp at the end because of a primer/adapter contamination

    Journal: NPJ Genomic Medicine

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection

    doi: 10.1038/s41525-017-0034-3

    Figure Lengend Snippet: Sequencing comparison in WGA amplified and non-amplified DNA samples. a Mutation detection comparison for REPLI-g WGA. REPLI-g amplified or non-amplified DNA from both fresh and fixed HCT116 cells were subjected to the CRC targeted NGS. The blue color highlights the true mutations detected, while the number inside each cell represents the MAF of the mutation. The light red color highlights the false mutations called. b Gene coverage comparison for WGA. The coverage was compared among the following samples: ① Fresh cells; ② Fixed cells; ③ Fresh cells + REPLI-g; ④ Fixed cells + REPLI-g. ⑤ Fixed cells+WGA4. The reads were aligned using BWA-MEM and coverage was computed using GATK’s DepthOfCoverage and in-house scripts. The percentage of bases covered by at least 10 reads with minimum base quality score Q30 (%_bases_above_10) was calculated using data obtained from DepthOfCoverage and in-house scripts and summarized in the table. (*) In the case of GenomePlex (WGA4), the plotted data was trimmed of 10 bp at the beginning and 40 bp at the end because of a primer/adapter contamination

    Article Snippet: For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Techniques: Sequencing, Whole Genome Amplification, Amplification, Mutagenesis, Next-Generation Sequencing

    Workflow optimization for biopsy genomic profiling, tissue and CTCs. Qiagen QIAamp Micro Kit and GeneRead DNAseq targeted panel sequencing were applied on all types of cell samples. This overall workflow performs well when DNA amount is sufficient, i.e., from whole blood and tissue biopsy. For rare cells, DNA input is insufficient for targeted NGS, leading to a bias and the need for an extra step of Whole Genome Amplification (WGA). Two WGA kits were evaluated (WGA4 from Sigma and REPLI-g from Qiagen) and obtained low coverage for fixed cells, while REPLI-g was identified as optimal for fresh rare cells and used for a final validation on patient CTCs

    Journal: NPJ Genomic Medicine

    Article Title: Workflow optimization of whole genome amplification and targeted panel sequencing for CTC mutation detection

    doi: 10.1038/s41525-017-0034-3

    Figure Lengend Snippet: Workflow optimization for biopsy genomic profiling, tissue and CTCs. Qiagen QIAamp Micro Kit and GeneRead DNAseq targeted panel sequencing were applied on all types of cell samples. This overall workflow performs well when DNA amount is sufficient, i.e., from whole blood and tissue biopsy. For rare cells, DNA input is insufficient for targeted NGS, leading to a bias and the need for an extra step of Whole Genome Amplification (WGA). Two WGA kits were evaluated (WGA4 from Sigma and REPLI-g from Qiagen) and obtained low coverage for fixed cells, while REPLI-g was identified as optimal for fresh rare cells and used for a final validation on patient CTCs

    Article Snippet: For REPLI-g Single Cell Kit: The standard protocol “Whole genome amplification from genomic DNA using the REPLI-g® Single Cell Kit” from Qiagen, as well as the protocol “increased volume” were both used.

    Techniques: Sequencing, Next-Generation Sequencing, Whole Genome Amplification