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PMC:1310901 / 8718-10201 JSONTXT 14 Projects

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Id Subject Object Predicate Lexical cue
T4350 0-42 Sentence denotes Methylation-specific restriction-PCR-assay
T64 0-42 Sentence denotes Methylation-specific restriction-PCR-assay
T4351 43-107 Sentence denotes DNA was extracted with a commercial kit (Qiagen) as recommended.
T65 43-107 Sentence denotes DNA was extracted with a commercial kit (Qiagen) as recommended.
T4352 108-351 Sentence denotes Since the restriction ability of several endonucleases is inhibited by methylation of their target sequence, we used methylation-sensitive enzymes HpaII and HaeII-isochizomer Bsp143II and Bsh1236I (MBI Fermentas, St Leon-Rot, Germany) (20,24).
T66 108-351 Sentence denotes Since the restriction ability of several endonucleases is inhibited by methylation of their target sequence, we used methylation-sensitive enzymes HpaII and HaeII-isochizomer Bsp143II and Bsh1236I (MBI Fermentas, St Leon-Rot, Germany) (20,24).
T4353 352-481 Sentence denotes As control the methylation-resistant enzyme MspI and an enzyme with no recognition site in the target promoter, EcoRI, were used.
T67 352-481 Sentence denotes As control the methylation-resistant enzyme MspI and an enzyme with no recognition site in the target promoter, EcoRI, were used.
T4354 482-605 Sentence denotes DNA (0.8 µg) was digested by 40 U the respective enzyme for 6 h and, to ensure complete cleavage, additional 20 U for 16 h.
T68 482-605 Sentence denotes DNA (0.8 µg) was digested by 40 U the respective enzyme for 6 h and, to ensure complete cleavage, additional 20 U for 16 h.
T4355 606-774 Sentence denotes Thereafter 100 ng of digested DNA was used to a PCR amplification of two fragments (F1 and F2) spanning part of the IRF-4 promoter (30) (GenBank U52683; see Figure 3A).
T69 606-774 Sentence denotes Thereafter 100 ng of digested DNA was used to a PCR amplification of two fragments (F1 and F2) spanning part of the IRF-4 promoter (30) (GenBank U52683; see Figure 3A).
T4356 775-820 Sentence denotes The sequences of the primers were F1-forward:
T70 775-820 Sentence denotes The sequences of the primers were F1-forward:
T4357 821-913 Sentence denotes 5′-TTGAGATGGAGTCTTGCTCTGT-3′, F1-reverse: ATCACTTCCAGACTTCAGTTCACCT-3′ (341 bp); F2-forward:
T71 821-913 Sentence denotes 5′-TTGAGATGGAGTCTTGCTCTGT-3′, F1-reverse: ATCACTTCCAGACTTCAGTTCACCT-3′ (341 bp); F2-forward:
T4358 914-958 Sentence denotes 5′-AAGGTGAACTGAAGTCTGGAAGTGA-3′, F2-reverse:
T72 914-958 Sentence denotes 5′-AAGGTGAACTGAAGTCTGGAAGTGA-3′, F2-reverse:
T4359 959-998 Sentence denotes 5′-CCAGGACCTCAGGAGGCCAGTCA-3′ (474 bp).
T73 959-998 Sentence denotes 5′-CCAGGACCTCAGGAGGCCAGTCA-3′ (474 bp).
T4360 999-1047 Sentence denotes The PCR conditions were described elsewhere (3).
T74 999-1047 Sentence denotes The PCR conditions were described elsewhere (3).
T4361 1048-1118 Sentence denotes PCR was performed with an annealing temperature of 62°C and 35 cycles.
T75 1048-1118 Sentence denotes PCR was performed with an annealing temperature of 62°C and 35 cycles.
T4362 1119-1319 Sentence denotes When DNA was methylated at specific sites, the sensitive enzymes were not able to digest the DNA and amplification took place; in case of no methylation, DNA was digested and no product was generated.
T76 1119-1319 Sentence denotes When DNA was methylated at specific sites, the sensitive enzymes were not able to digest the DNA and amplification took place; in case of no methylation, DNA was digested and no product was generated.
T4363 1320-1431 Sentence denotes The PCR products were electrophoresed on a 3% agarose gel, were stained with ethidium bromide and photographed.
T77 1320-1431 Sentence denotes The PCR products were electrophoresed on a 3% agarose gel, were stained with ethidium bromide and photographed.
T4364 1432-1483 Sentence denotes PCR products were verified by automated sequencing.
T78 1432-1483 Sentence denotes PCR products were verified by automated sequencing.