G&T-Seq
Genome and Transcriptome Sequencing
G&T-seq can separate and sequence gDNA 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 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 WGA method
- No 3ê-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 risk of sample loss or contamination
- Increased handling time
Reagents:
Illumina Library prep and Array Kit Selector
Reviews:
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
Saadatpour A., Lai S., Guo G. and Yuan G. C. Single-Cell Analysis in Cancer Genomics. Trends Genet. 2015;31:576-586
References:
Macaulay I. C., Haerty W., Kumar P., et al. G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nat Methods. 2015;12:519-522
Related
History: G&T-Seq
Revision by on 2017-06-21 09:15:21 - Show/Hide
Genome and Transcriptome Sequencing
G&T-seq can separate and sequence gDNA 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 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 WGA method
- No 3ê-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 risk of sample loss or contamination
- Increased handling time
Reagents:Illumina Library prep and Array Kit SelectorReviews: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;76:1305-1312Saadatpour A., Lai S., Guo G. and Yuan G. C. Single-Cell Analysis in Cancer Genomics. Trends Genet. 2015;31:576-586References:Macaulay I. C., Haerty W., Kumar P., et al. G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nat Methods. 2015;12:519-522Revision by sbrumpton on 2017-06-21 09:06:26 - Show/Hide
Genome and Transcriptome Sequencing
G&T-seq can separate and sequence gDNA 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 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 WGA method
- No 3ê-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 risk of sample loss or contamination
- Increased handling time
Reagents:Illumina Library prep and Array Kit SelectorReviews: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;76:1305-1312Saadatpour A., Lai S., Guo G. and Yuan G. C. Single-Cell Analysis in Cancer Genomics. Trends Genet. 2015;31:576-586References:Macaulay I. C., Haerty W., Kumar P., et al. G&T-seq: parallel sequencing of single-cell genomes and transcriptomes. Nat Methods. 2015;12:519-522