Ethidium Bromide → 13,14-dihydro-15-keto-PGE2

HRID 429031

反応式

HRID 429031 の反応式
S([O-])(OCCCCCCCCCCCC)(=O)=[O:2].[Na+].B(O)(O)O.C(N([CH2:39][C:40]([OH:42])=[O:41])CC(O)=O)CN(CC(O)=O)CC(O)=O.C(O)[C:44](N)([CH2:47][OH:48])[CH2:45][OH:46].CC[N+]1[C:66]([C:67]2[CH:68]=CC=CC=2)=[C:65]2[C:60]([CH:61]=[CH:62][C:63](N)=[CH:64]2)=[C:59]2[C:54]=1[CH:55]=[C:56](N)[CH:57]=[CH:58]2.[Br-]>>[CH3:54][CH2:55][CH2:56][CH2:57][CH2:58][C:59]([CH2:60][CH2:61][C@@H:62]1[C@@H:63]([CH2:64]/[CH:65]=[CH:66]\[CH2:67][CH2:68][CH2:39][C:40]([OH:42])=[O:41])[C:45](=[O:46])[CH2:44][C@H:47]1[OH:48])=[O:2]

参加物

4

反応物 3

HCID 14710Ethidium BromideHCID 3423265Sodium Lauryl SulfateHCID 212577252-Amino-2-(hydroxymethyl)propane-1,3-diol;2-(2-(bis(carboxymethyl)amino)ethyl-(carboxymethyl)amino)acetic acid;boric acid

生成物 1

HCID 528071113,14-dihydro-15-keto-PGE2

実験手順

After the integration reaction, each reaction was stop with sodium dodecyl sulfate and proteinase K at final concentrations of 1% and 1 mg/ml, respectively. The samples were further incubated at 37° C. for 2 hr and then subjected to phenol-chloroform (1:1) and ether extractions. The DNA was precipitated by ethanol. Aliquots of each sample were subjected to electrophoresis on 1 or 1.5% agarose gels which were dried and the radioactive products were quantitated by a Molecular Dynamics PhosphorImager. The dried gels were also exposed to X-ray films with or without an intensifying screen. For BglII restriction analysis of the M-2/pGEM recombinants, each 20 μl reaction mixture was digested by 12 units of Bglll for 2 hr to ensure complete digestion. The samples were subjected to electrophoresis on 1.5% agarose gels in a Tris-borate-EDTA buffer with 0.5 μg/ml of ethidium bromide for 13 hr at 100 volts. The gels were dried and analyzed as described above. Linear DNA fragments (Boehringer Mannheim), pGEM, and R35(Ref.2) were used as unlabeled molecular weight markers. Linear R35 (3-4 Kbp) was 5′ end labeled with γ-32P ATP and was used to identify the same size BglII linerarized M-2/pGEM recombinants which resulted from the concerted insertion of M-2 into pGEM.

後処理

  1. customAfter the integration reaction
  2. extractionsubjected to phenol-chloroform (1:1) and ether extractions
  3. customThe DNA was precipitated by ethanol
  4. customwere dried
  5. customFor BglII restriction analysis of the M-2/pGEM recombinants, each 20 μl reaction mixture
  6. waitwas digested by 12 units of Bglll for 2 hr
  7. customThe gels were dried

出典と根拠

ORDレコード
ord-489a15b24b424a29aad8285374435a1f
出典データセット
uspto-grants-2001_11
特許番号
US06316261B1

関連ノート

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