Methods for quenching a nuclease digestion of a target nucleic acid prior to downstream analysis of the target nucleic acid are disclosed herein. Particularly, methods for controlling the end point of a nuclease digestion prior to sequence analysis of a target nucleic acid is provided. Quenching of a nuclease digestion in the present disclosure employs at least one non-ionic or anionic denaturant combined with an optional reducing agent. The methods presented in this disclosure aids preserving the sample comprising the target nucleic acid or fragments thereof for long term storage and ensures that the effect of contaminating nucleases is eliminated during pretreatment step.

Described herein are methods of purifying polynucleotides, e.g., imRNA and oligonucleotides, e.g., probes, primers and siRNA, using monolithic columns with immobilized ligands coupled to the monolithic column. Also described are monolithic columns for purifying polynucleotides from a sample; and methods of preparing such columns.

Described herein are methods of purifying polynucleotides, e.g., imRNA and oligonucleotides, e.g., probes, primers and siRNA, using monolithic columns with immobilized ligands coupled to the monolithic column. Also described are monolithic columns for purifying polynucleotides from a sample; and methods of preparing such columns.

The present invention relates to the detection of a target nucleic acid sequence in a real-time manner using a target signal generating primer (TSG primer) having dual interactive labels. The present invention allows for both target amplification and signal amplification by introducing dual interactive labels into a primer used in PCR reactions, ensuring real-time target detection by PCR reactions without the use of complicated oligonucleotides. The present invention could be free from the troublesome matters and shortcomings associated with conventional real-time PCR methods. The present invention allows for successful real-time target detection by using only a labeled primer. Also, the present invention can obtain strong signals indicative of the presence of target nucleic acid sequences in both a liquid phase and solid phase.

The present invention relates to the detection of a target nucleic acid sequence in a real-time manner using a target signal generating primer (TSG primer) having dual interactive labels. The present invention allows for both target amplification and signal amplification by introducing dual interactive labels into a primer used in PCR reactions, ensuring real-time target detection by PCR reactions without the use of complicated oligonucleotides. The present invention could be free from the troublesome matters and shortcomings associated with conventional real-time PCR methods. The present invention allows for successful real-time target detection by using only a labeled primer. Also, the present invention can obtain strong signals indicative of the presence of target nucleic acid sequences in both a liquid phase and solid phase.

Provided herein includes a method for characterizing a target cell-free nucleic acid (cfNA) molecule present in a biological sample such as a urine sample. It comprises isolating total cfNAs from the biological sample without prior preprocessing such as centrifugation to remove cell debris, and characterizing the target cfNA molecule based on the isolated total cfNAs. When the target cfNA is a low molecular weight (LMW) molecule, the method additionally comprises a fractionation step to obtain LMW nucleic acids from the total cfNAs before characterization. The method can detect significantly more copies of the target cfNA molecule compared with existing methods which typically discard the cell debris from the biological sample. Another method is also provided, which substantially recovers cfNAs from the usually discarded cell debris, thus also capable of detecting significantly more copies of the target cfNA molecule.

Provided herein includes a method for characterizing a target cell-free nucleic acid (cfNA) molecule present in a biological sample such as a urine sample. It comprises isolating total cfNAs from the biological sample without prior preprocessing such as centrifugation to remove cell debris, and characterizing the target cfNA molecule based on the isolated total cfNAs. When the target cfNA is a low molecular weight (LMW) molecule, the method additionally comprises a fractionation step to obtain LMW nucleic acids from the total cfNAs before characterization. The method can detect significantly more copies of the target cfNA molecule compared with existing methods which typically discard the cell debris from the biological sample. Another method is also provided, which substantially recovers cfNAs from the usually discarded cell debris, thus also capable of detecting significantly more copies of the target cfNA molecule.

The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.

The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.

Methods for degrading contaminant nucleic acid. The methods use combinations of metal ions and peroxide ions to produce a variety of oxidative species that degrade nucleic acid. Methods of the invention are useful for decontaminating laboratory equipment or solutions. After the equipment or solutions have been decontaminated, the metal ion and peroxide ion solution can be deactivated by raising the temperature to dissociate the peroxide or by binding the metal ions, e.g., with a chelating agent.