Ethylenediaminetetraacetic Acid → 2'-Deoxycytidine-5'-Monophosphate

HRID 1590311

Reaktionsgleichung

Reaktionsgleichung von HRID 1590311
[As](C)(C)(=O)[O-].C(S)[C@@H](O)[C@H](O)CS.[P:14]([O:26][CH2:27][C@H:28]1[O:32][C@@H:31]([N:33]2[CH:40]=[CH:39][C:37]([NH2:38])=[N:36][C:34]2=[O:35])[CH2:30][C@@H:29]1[OH:41])([O:17]P(OP(O)(O)=O)(O)=O)(=[O:16])[OH:15].C(N(CC(O)=O)CC(O)=O)CN(CC(O)=O)CC(O)=O>O>[CH2:30]1[C@H:31]([N:33]2[C:34](=[O:35])[N:36]=[C:37]([NH2:38])[CH:39]=[CH:40]2)[O:32][C@H:28]([CH2:27][O:26][P:14]([OH:16])([OH:17])=[O:15])[C@H:29]1[OH:41]

Komponenten

6

Edukt 4

HCID 6049Ethylenediaminetetraacetic AcidHCID 65091dCTPHCID 167250CacodylateHCID 446094D-1,4-Dithiothreitol

Produkt 1

HCID 139452'-Deoxycytidine-5'-Monophosphate

Reagenzien, Katalysatoren, Lösungsmittel 1

HCID 962WaterLösungsmittel

Versuchsvorschrift

Double-stranded cDNA was homopolymer tailed essentially by the method of R. Roychoudhury and R. Wu, Meth. Enz., 65, pp. 43-62 (1980). The following were added together, in order: 3 μl dCTP32 (>800 Ci/mmol), 20 μl 10 X cacodylate buffer (1.4 mM potassium cacodylate, 0.3M Tris-hydrochloride, pH 7.6), 4 μl 5 mM DTT, 3 μl 10 mM dCTP, 2 μl 100 mM CoCl2, 12 units terminal deoxynucleotide transferase (PL Biochemicals; at least 8 units/pmol 3' end) in 1.5 μl, H2O to 200 μl final volume. The reaction was carried out at 12° for 1.5 min then stopped by adding 10 μl 0.5M EDTA This reaction resulted in an average of 15 dCMP residues added per 3' end of dsDNA, the optimal number for annealing and transformation (Peacock et al., Bioch. Biophys. Acta, 655, pp. 243-50 (1981)

Aufarbeitung

  1. additionThe following were added together, in order

Quellen und Nachweise

ORD-Eintrag
ord-0c084c41e25d412587e8f744f87f5241
Quelldatensatz
uspto-grants-1993_04
Patentnummer
US05202430

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