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

HRID 1590311

반응식

HRID 1590311 반응식
[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]

참가 물질

6

반응물 4

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

생성물 1

HCID 139452'-Deoxycytidine-5'-Monophosphate

시약·촉매·용매 1

HCID 962Water용매

실험 절차

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)

후처리

  1. additionThe following were added together, in order

출처 및 근거

ORD 기록
ord-0c084c41e25d412587e8f744f87f5241
출처 데이터셋
uspto-grants-1993_04
특허 번호
US05202430

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