A polynucleotide sequencing method includes a wash step that employs a composition including a polymerase. The composition may also include a plurality of nucleotides. The composition may be configured to prevent the polymerase from incorporating one of the plurality of nucleotides into a copy polynucleotide strand. The composition may be substantially free of Mg.sup.2+.

A polynucleotide sequencing method includes a wash step that employs a composition including a polymerase. The composition may also include a plurality of nucleotides. The composition may be configured to prevent the polymerase from incorporating one of the plurality of nucleotides into a copy polynucleotide strand. The composition may be substantially free of Mg.sup.2+.

Methods of assaying cells of a cellular sample for the presence of a target nucleic acid are provided. Aspects of the methods include evaluating a cellular sample that has been contacted with a nuclease inhibitor for the presence of a target nucleic acid. Also provided are devices and kits that find use in practicing the methods described herein.

Methods of assaying cells of a cellular sample for the presence of a target nucleic acid are provided. Aspects of the methods include evaluating a cellular sample that has been contacted with a nuclease inhibitor for the presence of a target nucleic acid. Also provided are devices and kits that find use in practicing the methods described herein.

A nucleic acid amplification reaction method includes subjecting a reaction mixture containing a nucleic acid amplification reaction reagent to be used for amplifying a nucleic acid to a thermal cycle for amplifying the nucleic acid, wherein in the thermal cycle, a heating time for an annealing reaction and an elongation reaction is 1 sec or more and 10 sec or less, the nucleic acid amplification reaction reagent contains a forward primer, a reverse primer, a polymerase, and a fluorescently labeled probe, the concentration of the forward primer is 0.4 μM or more and 3.2 μM or less, the concentration of the reverse primer is 0.4 μM or more and 3.2 μM or less, the amount of the polymerase is 0.5 U or more and 4 U or less, and the concentration of the fluorescently labeled probe is 0.15 μM or more and 1.2 μM or less.

A nucleic acid amplification reaction method includes subjecting a reaction mixture containing a nucleic acid amplification reaction reagent to be used for amplifying a nucleic acid to a thermal cycle for amplifying the nucleic acid, wherein in the thermal cycle, a heating time for an annealing reaction and an elongation reaction is 1 sec or more and 10 sec or less, the nucleic acid amplification reaction reagent contains a forward primer, a reverse primer, a polymerase, and a fluorescently labeled probe, the concentration of the forward primer is 0.4 μM or more and 3.2 μM or less, the concentration of the reverse primer is 0.4 μM or more and 3.2 μM or less, the amount of the polymerase is 0.5 U or more and 4 U or less, and the concentration of the fluorescently labeled probe is 0.15 μM or more and 1.2 μM or less.

The present invention relates to a method of screening for the presence of methylated DNA in a biological sample by using multiple methylation-sensitive restriction endonucleases. More particularly, the present invention relates to a method of quantitatively screening for the level of one or more methylated genes of interest without the requirement that an undigested internal reference sample is used as a point of reference against which relative quantification is calculated. The present invention is useful in a range of applications including, but not limited to, providing a simpler and more accurate means to determine DNA methylation status, such as in the context of diagnosing or monitoring conditions characterised by changes to DNA methylation.

The present invention relates to a method of screening for the presence of methylated DNA in a biological sample by using multiple methylation-sensitive restriction endonucleases. More particularly, the present invention relates to a method of quantitatively screening for the level of one or more methylated genes of interest without the requirement that an undigested internal reference sample is used as a point of reference against which relative quantification is calculated. The present invention is useful in a range of applications including, but not limited to, providing a simpler and more accurate means to determine DNA methylation status, such as in the context of diagnosing or monitoring conditions characterised by changes to DNA methylation.

The present disclosure provides methods and kits for ligation-coupled PCR. Methods for performing ligation and PCR and, optionally, enzymatic digestion in a single closed tube are provided. Methods and kits for splint-mediated primer assembly and ligation-coupled PCR are also provided.

The present disclosure provides methods and kits for ligation-coupled PCR. Methods for performing ligation and PCR and, optionally, enzymatic digestion in a single closed tube are provided. Methods and kits for splint-mediated primer assembly and ligation-coupled PCR are also provided.