ライフサイエンスコーパス: ddNTP

1 g and rate constants were measured for 2',3'-ddNTPs as well as for several other 3'-substituted termi

2 for phosphodiester bond formation for 2',3'-ddNTPs were 200-3000-fold lower than for dNTPs.

3 did not eliminate selectivity against 2',3'-ddNTPs.

4 d complex is not formed in the presence of a ddNTP or rNTP.

5 Wild-type Pfu-Pol required a ddNTP:dNTP ratio of 30:1; values of 5:1 (Q472H), 1:3 (L4

6 Primers that do not acquire ddNTP are used to capture and amplify the unique target

7 ryl transfer, whereas discrimination against ddNTPs was almost exclusively due to a slower rate of ph

8 matic decrease in the discrimination against ddNTPs, while mutations in Tyr766 and Glu710 had a small

9 ddATP is the least efficient substrate among ddNTPs examined.

10 Both dNTP and ddNTP are substrates of the polymerase in the presence o

11 ing the gapped DNA with a blocked primer and ddNTP.

12 arameters for the incorporation of dNTPs and ddNTPs were determined for wild-type Klenow fragment and

13 nt roles in distinguishing between dNTPs and ddNTPs.

14 are related to the engineered polymerase and ddNTPs which are intrinsic to any sequencing-by-synthesi

15 was extended by DNA polymerase with a biotin-ddNTP.

16 The use of CFET-labeled primers and biotin-ddNTPs coupled with the specificity of DNA polymerase in

17 able biotinylated dideoxynucleotides (biotin-ddNTPs) in single base extension for multiplex genotypin

18 and biotinylated dideoxynucleotides (biotin-ddNTPs).

19 We selected four biotin-ddNTPs with distinct molecular weights to generate exten

20 in the DNA template are extended with biotin-ddNTPs by DNA polymerase to generate 3'-biotinylated DNA

21 simultaneously in a single tube with biotin-ddNTPs to generate 3(')-biotinylated DNA products, which

22 ligonucleotide primers was mixed with biotin-ddNTPs, DNA polymerase and the DNA templates containing

23 s, metal ions, and dNTPs and is inhibited by ddNTPs or chain-terminated DNA.

24 luorophore on the DNA products terminated by ddNTPs.

25 When the Watson-Crick complementary ddNTP is added to the SBE reaction, the primer is extend

26 ifference compared to that with conventional ddNTPs.

27 porate the correct dNTP, but not the correct ddNTP, was also observed in the presence of Mg(2+).

28 inhibition of HIV-1 RT as the corresponding ddNTPs.

29 ay detects the unreacted dideoxynucleotides (ddNTPs) remaining or surviving in solution following a S

30 y cleavable biotinylated dideoxynucleotides, ddNTPs-N(3)-biotin, for the DNA polymerase extension rea

31 from the structures of other polymerase-DNA-ddNTP complexes in that the 3'-terminus of the primer ha

32 nd dideoxynucleoside-5'-triphosphate (RT-DNA-ddNTP), utilizing the ddNTPs ddATP, betaFddATP, and alph

33 ses, we propose active site models for dNTP, ddNTP, and acyNTP selection by hyperthermophilic archaea

34 tant derivatives that showed changes in dNTP/ddNTP discrimination.

35 By varying the dNTP/ddNTP mixtures used, the assay could also be directed to

36 Mn(2+), 3D(pol) efficiently utilized dNTPs, ddNTPs, and incorrect NTPs.

37 elity and kinetic parameters for DNA and dye-ddNTPs.

38 hibit similar K(m) and V(max) values for dye-ddNTPs in single-base extension assays, the archaeal mut

39 the SNaPshot kit containing four fluorescent-ddNTPs using the CE technology, and explored various str

40 Therefore, the increased selectivity for ddNTPs is likely to arise from two factors: (i) small ov

41 t yield the desired level of selectivity for ddNTPs.

42 etermined the crystal structures of all four ddNTP-trapped closed ternary complexes of the large frag

43 hesized four 3'-O-azidomethyl-dNTPs and four ddNTP-azidolinker-fluorophores for the hybrid SBS.

44 involves a simple analysis of the same four ddNTP analytes, regardless of the SNP being investigated

45 M was highly discriminatory against all four ddNTPs but was able to incorporate rNTPs as efficiently

46 itivity of the wild-type MuLV RT to all four ddNTPs remained unchanged by mutation of V223 to Met or

47 The resistance to all four ddNTPs, however, persisted in the wild type and Y222 mut

48 and screening mutant libraries for improved ddNTP incorporation.

49 wo mutations, P410L and A485T, that improved ddNTP uptake, suggesting the contribution of P410 and A4

50 L/A485T) has an additive effect in improving ddNTP incorporation by a total of 250-fold.

51 yrococcus furiosus with the aim of improving ddNTP utilisation.

52 eins, suggesting no participation of K152 in ddNTP recognition.

53 97) gave rise to variants which incorporated ddNTPs better than the wild type, allowing sequencing re

54 The determination of Ki(ddNTP)/K(m)(dNTP) ratios showed that AZTTP, D4TTP, and d

55 nsferase addition of a fluorescently labeled ddNTP.

56 ncing reactions to be carried out at lowered ddNTP:dNTP ratios.

57 We examined the molecular basis of ddNTP selectivity in archaeal family B DNA polymerases b

58 molecular mechanism of RT discrimination of ddNTPs from natural substrates.

59 own preference to bind the incoming dNTP (or ddNTP) with a north conformation at the polymerase site,

60 ry to the mutated nucleotide and three other ddNTPs and were optimized to quantify levels of a mutati

61 condary roles in the selection of dNTPs over ddNTPs.In order to understand the interactions in the en

62 Ps) by Vent DNA polymerase was enhanced over ddNTPs via a 50-fold increase in phosphoryl transfer rat

63 nd variant incorporated both dNTP-ONH(2)sand ddNTPs faithfully and efficiently, supporting extension-

64 previously shown to be resistant to several ddNTPs and to phosphonoformic acid.

65 -flanking region of the gene, using SNaPshot ddNTP primer extension and capillary electrophoresis.

66 yped for polymorphisms of MIF using SNaPshot ddNTP primer extension, or by a fluorescently labeled pr

67 cleotide triphosphate binding site such that ddNTPs become favoured; (ii) interference with a conform

68 The ddNTP resistance patterns were unchanged for all muteins

69 sition the dNTP, but specificity against the ddNTP is lost because the phenolic OH can compensate for

70 NP, without any labeling requirements of the ddNTPs or oligonucleotide primers.

71 ble linker to attach the fluorophores to the ddNTPs, we synthesized four 3'-O-azidomethyl-dNTPs and f

72 '-triphosphate (RT-DNA-ddNTP), utilizing the ddNTPs ddATP, betaFddATP, and alphaFddATP, explain the e

73 re from the DNA products terminated with the ddNTPs, the polymerase reaction reinitiates to continue

74 by mini-sequencing in the presence of three ddNTPs and the fourth nucleotide in the deoxy form.

75 porated 10 times faster than the other three ddNTPs), and (ii) these enzymes show uneven band-intensi

76 gesting the contribution of P410 and A485 to ddNTP/dNTP selectivity in archaeal DNA polymerases.

77 lymerases, while the contribution of P410 to ddNTP/dNTP selectivity has not been reported.

78 151N mutant was only moderately resistant to ddNTPs but exhibited a higher level of discrimination ag

79 d Q151N) and examined their sensitivities to ddNTPs and their ability to discriminate against rNTPs v

80 the Y416F mutant was more permissive toward ddNTP vs rNTP utilization than either the Y416A mutant o

81 the four dideoxynucleoside 5' triphosphates (ddNTPs) at widely different rates during sequencing (ddG

82 used 2',3'-dideoxynucleoside triphosphates (ddNTPs), dNTP-ONH(2)s terminate primer extension.

83 on of 2',3'-dideoxynucleotide triphosphates (ddNTPs) indicating very different substrate selectivitie

84 Unlike ddNTPs, however, primer extension can be resumed by clea

85 l-I family which are able to efficiently use ddNTPs have demonstrated a much improved performance whe

86 oligonucleotides by extension reaction using ddNTPs and a double stranded DNA template generated from

87 rved cross-resistance of HIV-1 RT to various ddNTPs should reflect the alteration of RT's substrate r

88 s with about the same preference as dNTPs vs ddNTPs.