- It is recommended to perform the PCR reaction at half scale,
saving half of the template in case more or fewer PCR cycles are
needed. - DNA libraries may be stored at 20 C until sequencing.
References
- Desrosiers R, Friderici K, Rottman F (1974)
Identification of methylated nucleosides in mes-
senger RNA from Novikoff hepatoma cells.
Proc Natl Acad Sci U S A 71:3971–3975.
doi:10.1073/pnas.71.10.3971 - Wang X, Lu Z, Gomez A et al (2014) N6-
methyladenosine-dependent regulation of
messenger RNA stability. Nature
505:117–120. doi:10.1038/nature12730 - Wang X, Zhao BS, Roundtree IA et al (2015)
N^6 -methyladenosine modulates messenger
RNA translation efficiency. Cell 161:
1388–1399. doi:10.1016/j.cell.2015.05.014 - Bokar JA (2005) The biosynthesis and func-
tional roles of methylated nucleosides in
eukaryotic mRNA. In: Grosjean H (ed) Fine-
tuning of RNA functions by modification and
editing, vol 12. Springer, Berlin, pp 141–177.
doi:10.1007/b106365 - Dominissini D, Moshitch-Moshkovitz S,
Schwartz S et al (2012) Topology of the
human and mouse m6A RNA methylomes
revealed by m6A-seq. Nature 485:201–206.
doi:10.1038/nature11112 - Meyer KD, Saletore Y, Zumbo P et al (2012)
Comprehensive analysis of mRNA methylation
reveals enrichment in 3^0 UTRs and near stop
codons. Cell 149:1635–1646. doi:10.1016/j.
cell.2012.05.003 - Wei C, Moss B (1977) Nucleotide sequences at
the N6-methyladenosine sites of HeLa cell
messenger ribonucleic acid. Biochemistry
16:1672–1676. doi:10.1021/bi00627a023
8. Chen K, Lu Z, Wang X et al (2014) High-
resolution N(6)-methyladenosine (m(6) A)
map using photo-crosslinking-assisted m(6) A
sequencing. Angew Chem Int Ed Engl:1–5.
doi:10.1002/anie.201410647
9. Linder B, Grozhik A V, Olarerin-George AO
et al (2015) Single-nucleotide-resolution
mapping of m6A and m6Am throughout the
transcriptome. Nat Methods 12:767–772.
doi:10.1038/nmeth.3453 - Molinie B, Wang J, Lim KS, et al (2016) m 6 A-
LAIC-seq reveals the census and complexity of
the m 6 A epitranscriptome. doi:10.1038/
nmeth.3898 - Shi H, He C (2016) A glance at N6-
methyladenosine in transcript isoforms. Nat
Methods 13:624–625. doi:10.1038/nmeth.
3928 - Dominissini D, Moshitch-Moshkovitz S,
Salmon-Divon M et al (2013) Transcriptome-
wide mapping of N(6)-methyladenosine by m
(6)A-seq based on immunocapturing and mas-
sively parallel sequencing. Nat Protoc
8:176–189. doi:10.1038/nprot.2012.148 - Dominissini D, Moshitch-Moshkovitz S,
Amariglio N, Rechavi G (2015)
Transcriptome-wide mapping of N6-
methyladenosine by m6A-seq, 1st ed. RNA
Modif. doi:10.1016/bs.mie.2015.03.001 - Schwartz S, Agarwala SD, Mumbach MR et al
(2013) High-resolution mapping reveals a con-
served, widespread, dynamic mRNA methyla-
tion program in yeast meiosis. Cell 155:
1409–1421. doi:10.1016/j.cell.2013.10.047
Identifying the m^6 A Methylome 57