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Each MSP reaction incorporated approximately 20 ng of bisulfite-treated DNA as a template.
The second reaction incorporated the products of the first reaction, along with NewPLFsh (5′-CTATCAGCTTTTGATGTTAG-3′) and NewRevsh (5′-CCTTAACTTTCGTTCTTG-3′) primers.
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RT reactions incorporated Superscript II Reverse Transcriptase (Invitrogen), 0.5 μg of oligo (dT 17 and 0.5 μl of Prime Recombinant Ribonuclease Inhibitor (Eppendorf).
Reverse transcription reactions incorporated Superscript II Reverse Transcriptase (Gibco), 0.5 μg Oligo (dT 12−18 and 0.5 μl Prime Recombinant Ribonuclease Inhibitor (Eppendorf).
Redox reaction of incorporated Pr (Pr4+ to Pr3+) hardly occurs under both reductive and oxidative conditions.
Additionally, pyridinium cations generated during catalytic reaction and incorporated into the formed alkali bronze phases stabilise these structures.
Excellent regioselectivities were observed for the fluorination reaction (fluorine incorporated ortho to the R group; Scheme 2 b); however, the addition of the SCF3 and CF3 groups tended to produce mixtures.
After the crosslinking reaction, the incorporated azide-handles were biotinylated via the CuAAC by incubating the (crosslinked) HspB6 (1.7 μM) with biotin-alkyne (0.1 mM) for 1 h at 4°C in the presence of CuSO4, sodium ascorbate and triazole ligand (1 mM each).
As monomers are truncated during chemical reactions while incorporated in the polymer, s2m needs to delete the lost atoms before searching for them in the polymer.
We suggest that nonefficient reactions were incorporated by neutral processes.
Briefly, total RNA was reverse transcribed using an anchored oligo d(T 20 primer in cDNA synthesis reactions that incorporated aminoallyl- and aminohexyl-modified nucleotides.
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