T-RHEX-RNAseq - a tagmentation-based, rRNA blocked, random hexamer primed RNAseq method for generating stranded RNAseq libraries directly from very low numbers of lysed cells.
Expression profiling
RNA purification free
Random hexamer priming
Stranded RNAseq
Tagmentation
Journal
BMC genomics
ISSN: 1471-2164
Titre abrégé: BMC Genomics
Pays: England
ID NLM: 100965258
Informations de publication
Date de publication:
17 Apr 2023
17 Apr 2023
Historique:
received:
25
10
2022
accepted:
28
03
2023
medline:
19
4
2023
entrez:
18
4
2023
pubmed:
19
4
2023
Statut:
epublish
Résumé
RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming - to allow analysis of a broader range of transcripts - remains challenging. We here describe a tagmentation-based, rRNA blocked, random hexamer primed RNAseq approach (T-RHEX-RNAseq) for generating stranded RNAseq libraries from very low numbers of FACS sorted cells without RNA purification steps. T-RHEX-RNAseq provides an easy-to-use, time efficient and automation compatible method for generating stranded RNAseq libraries from rare cells.
Sections du résumé
BACKGROUND
BACKGROUND
RNA sequencing has become the mainstay for studies of gene expression. Still, analysis of rare cells with random hexamer priming - to allow analysis of a broader range of transcripts - remains challenging.
RESULTS
RESULTS
We here describe a tagmentation-based, rRNA blocked, random hexamer primed RNAseq approach (T-RHEX-RNAseq) for generating stranded RNAseq libraries from very low numbers of FACS sorted cells without RNA purification steps.
CONCLUSION
CONCLUSIONS
T-RHEX-RNAseq provides an easy-to-use, time efficient and automation compatible method for generating stranded RNAseq libraries from rare cells.
Identifiants
pubmed: 37069502
doi: 10.1186/s12864-023-09279-4
pii: 10.1186/s12864-023-09279-4
pmc: PMC10111750
doi:
Substances chimiques
RNA, Ribosomal
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
205Informations de copyright
© 2023. The Author(s).
Références
Nat Protoc. 2014 Jan;9(1):171-81
pubmed: 24385147
Nat Methods. 2017 Mar;14(3):267-270
pubmed: 28092691
Nat Commun. 2018 Feb 12;9(1):619
pubmed: 29434199
Nat Biotechnol. 2020 Mar;38(3):276-278
pubmed: 32055031
Nature. 2009 Jan 8;457(7226):191-5
pubmed: 19011611
Nat Biotechnol. 2022 Oct;40(10):1452-1457
pubmed: 35637418
BMC Med Genomics. 2019 May 22;12(1):66
pubmed: 31118097
Bioinformatics. 2016 Oct 1;32(19):3047-8
pubmed: 27312411
Stem Cell Reports. 2022 Jul 12;17(7):1546-1560
pubmed: 35714596
Proc Natl Acad Sci U S A. 2021 Dec 21;118(51):
pubmed: 34911763
Nat Genet. 2014 Dec;46(12):1343-9
pubmed: 25326703
Science. 2008 Dec 19;322(5909):1855-7
pubmed: 19056939
Sci Rep. 2019 May 8;9(1):7091
pubmed: 31068632
Science. 2007 Jun 8;316(5830):1484-8
pubmed: 17510325
Genome Biol. 2015 Jul 23;16:148
pubmed: 26201400
Curr Protoc Mol Biol. 2015 Jan 05;109:21.29.1-21.29.9
pubmed: 25559105
Nat Commun. 2022 Jul 26;13(1):4057
pubmed: 35882841
Genome Biol. 2010;11(12):R119
pubmed: 21143862
Nat Biotechnol. 2022 Dec;40(12):1780-1793
pubmed: 35760914
Nat Rev Genet. 2019 Nov;20(11):631-656
pubmed: 31341269
BMC Genomics. 2019 Jan 18;20(1):59
pubmed: 30658577
Nat Methods. 2015 Oct;12(10):963-965
pubmed: 26280331
Genome Res. 2002 Jun;12(6):996-1006
pubmed: 12045153
RNA. 2020 Oct;26(10):1481-1488
pubmed: 32503920
Nat Methods. 2013 Jul;10(7):623-9
pubmed: 23685885
Nat Biotechnol. 2020 Jun;38(6):708-714
pubmed: 32518404
Sci Rep. 2019 May 17;9(1):7550
pubmed: 31101892
Genome Res. 2012 Jan;22(1):134-41
pubmed: 22128135
Nucleic Acids Res. 2009 Oct;37(18):e123
pubmed: 19620212
Proc Natl Acad Sci U S A. 1999 Aug 3;96(16):9045-50
pubmed: 10430892
Sci Rep. 2022 Jan 12;12(1):621
pubmed: 35022475
J Exp Med. 2022 Feb 7;219(2):
pubmed: 34919144