G&T-Seq
Genome and Transcriptome Sequencing
G&T-Seq can separate and sequence genomic DNA and full-length mRNA from single cells (Macaulay et al., 2015). In this method, single cells are isolated and lysed. RNA is captured using biotinylated oligo(dT) capture primers and separated from DNA using streptavidin-coated magnetic beads. Smart-seq2 is used to amplify captured RNA on the bead, while multiple displacement amplification (MDA) is used to amplify DNA. After sequencing, integrating the DNA and RNA sequences provides insights into the gene expression profiles of single cells.
Advantages:
- Compatible with any whole-genome amplification method
- No 3′-end bias in sequence reads because full-length transcripts are captured
- Because DNA and RNA are physically separated and amplified independently, there is no need to mask coding sequences during analysis
Disadvantages:
- Physical separation of DNA and RNA can increase the risk of sample loss or contamination
- Physical separation of DNA and RNA increases handling time
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
Bock C., Farlik M. and Sheffield N. C. Multi-Omics of Single Cells: Strategies and Applications. Trends Biotechnol. 2016;34:605-608
Clark S. J., Lee H. J., Smallwood S. A., Kelsey G. and Reik W. Single-cell epigenomics: powerful new methods for understanding gene regulation and cell identity. Genome Biol. 2016;17:72
Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X., et al. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312
Vieira Braga F. A., Teichmann S. A. and Chen X. Genetics and immunity in the era of single-cell genomics. Hum Mol Genet. 2016;25:R141-R148
Lu L., Lv B., Huang K., Xue Z., Zhu X. and Fan G. Recent advances in preimplantation genetic diagnosis and screening. J Assist Reprod Genet. 2016;33:1129-1134
References:
Angermueller C., Clark S. J., Lee H. J., et al. Parallel single-cell sequencing links transcriptional and epigenetic heterogeneity. Nat Methods. 2016;13:229-232
Related
History: G&T-Seq
Revision by on 2017-06-21 07:50:24 - Show/Hide
Genome and Transcriptome Sequencing
G&T-Seq can separate and sequence genomic DNA and full-length mRNA from single cells (Macaulay et al., 2015). In this method, single cells are isolated and lysed. RNA is captured using biotinylated oligo(dT) capture primers and separated from DNA using streptavidin-coated magnetic beads. Smart-seq2 is used to amplify captured RNA on the bead, while multiple displacement amplification (MDA) is used to amplify DNA. After sequencing, integrating the DNA and RNA sequences provides insights into the gene expression profiles of single cells.
Advantages:- Compatible with any whole-genome amplification method
- No 3'-end bias in sequence reads because full-length transcripts are captured
- Because DNA and RNA are physically separated and amplified independently, there is no need to mask coding sequences during analysis
Disadvantages:- Physical separation of DNA and RNA can increase the risk of sample loss or contamination
- Physical separation of DNA and RNA increases handling time
Reagents:Illumina Library prep and Array Kit SelectorReviews:Bock C., Farlik M. and Sheffield N. C. Multi-Omics of Single Cells: Strategies and Applications. Trends Biotechnol. 2016;34:605-608Clark S. J., Lee H. J., Smallwood S. A., Kelsey G. and Reik W. Single-cell epigenomics: powerful new methods for understanding gene regulation and cell identity. Genome Biol. 2016;17:72Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X., et al. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312Vieira Braga F. A., Teichmann S. A. and Chen X. Genetics and immunity in the era of single-cell genomics. Hum Mol Genet. 2016;25:R141-R148Lu L., Lv B., Huang K., Xue Z., Zhu X. and Fan G. Recent advances in preimplantation genetic diagnosis and screening. J Assist Reprod Genet. 2016;33:1129-1134References:Angermueller C., Clark S. J., Lee H. J., et al. Parallel single-cell sequencing links transcriptional and epigenetic heterogeneity. Nat Methods. 2016;13:229-232Revision by sbrumpton on 2017-06-21 07:50:20 - Show/Hide
Genome and Transcriptome Sequencing
G&T-Seq can separate and sequence genomic DNA and full-length mRNA from single cells (Macaulay et al., 2015). In this method, single cells are isolated and lysed. RNA is captured using biotinylated oligo(dT) capture primers and separated from DNA using streptavidin-coated magnetic beads. Smart-seq2 is used to amplify captured RNA on the bead, while multiple displacement amplification (MDA) is used to amplify DNA. After sequencing, integrating the DNA and RNA sequences provides insights into the gene expression profiles of single cells.
Advantages:- Compatible with any whole-genome amplification method
- No 3'-end bias in sequence reads because full-length transcripts are captured
- Because DNA and RNA are physically separated and amplified independently, there is no need to mask coding sequences during analysis
Disadvantages:- Physical separation of DNA and RNA can increase the risk of sample loss or contamination
- Physical separation of DNA and RNA increases handling time
Reagents:Illumina Library prep and Array Kit SelectorReviews:Bock C., Farlik M. and Sheffield N. C. Multi-Omics of Single Cells: Strategies and Applications. Trends Biotechnol. 2016;34:605-608Clark S. J., Lee H. J., Smallwood S. A., Kelsey G. and Reik W. Single-cell epigenomics: powerful new methods for understanding gene regulation and cell identity. Genome Biol. 2016;17:72Zhang X., Marjani S. L., Hu Z., Weissman S. M., Pan X., et al. Single-Cell Sequencing for Precise Cancer Research: Progress and Prospects. Cancer Res. 2016;76:1305-1312Vieira Braga F. A., Teichmann S. A. and Chen X. Genetics and immunity in the era of single-cell genomics. Hum Mol Genet. 2016;25:R141-R148Lu L., Lv B., Huang K., Xue Z., Zhu X. and Fan G. Recent advances in preimplantation genetic diagnosis and screening. J Assist Reprod Genet. 2016;33:1129-1134References:Angermueller C., Clark S. J., Lee H. J., et al. Parallel single-cell sequencing links transcriptional and epigenetic heterogeneity. Nat Methods. 2016;13:229-232