Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesized. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

Abstract: This specification relates to the field of molecular biology and provides novel methods and reagents for preserving and protecting the ribonucleic acid (RNA) content of samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.

Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesised. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

Abstract: The present invention relates to nuclease resistant nucleic acids in general and ribonuclease resistant RNAs in particular. Methods of making and using such nucleic acids are disclosed.

Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA trancription and in various other enzymatic reactions in which a polynucleotide is synthesised. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.

Abstract: The present invention is directed to compositions and methods for removal of nucleic acid probes from sample nucleic acids, particularly when the sample nucleic acids are attached to a solid support. The invention also concerns methods of stripping and reusing nucleic acid blots.

Abstract: The present invention is a general method for inactivating or inhibiting ribonucleases. Ribonucleases are treated with a reducing agent and heat. RNA samples contaminated with ribonuclease may be treated with this method to protect them from degradation. The RNA may then be used directly in a variety of enzymatic reactions and molecular biology techniques. This method may also be applied to a variety of molecular biology reagents which may be contaminated with ribonuclease to protect an RNA from being degraded when incubated with the reagent.

Abstract: This specification relates to the field of molecular biology and provides novel methods and reagents for preserving and protecting the ribonucleic acid (RNA) content of samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.

Abstract: This specification relates to the field of molecular biology and provides a novel method and reagent for preserving and protecting the ribonucleic acid (RNA) content of tissue samples from degradation prior to RNA isolation. This preservation may be accomplished without ultra-low temperature storage or disruption of the tissue.

Abstract: The invention relates to modified oligonucleotide primers used to adjust the amplification efficiency of an abundant target without affecting the amplification of other targets in a DNA synthesis reaction. The invention may be used in PCR.TM. or any other primer dependent DNA transcription technology.

Abstract: The present invention relates to nuclease resistant nucleic acids in general and ribonuclease resistant RNAs in particular. Methods of making and using such nucleic acids are disclosed.

Abstract: The present invention is directed to the process of creating a recombinant nucleic acid standard which is resistant to ribonuclease digestion and is non-infectious. A single strand of recombinant nucleic acid is encapsidated by bacteriophage proteins. The recombinant nucleic acid is a hybrid sequence encoding bacteriophage proteins and a specific non-bacteriophage sequence. A non-bacteriophage RNA sequence can be used as an RNA standard to help quantify the number of RNA molecules in an unknown sample. The recombinant RNA in its packaged form is highly resistant to ribonucleases, insuring that the RNA standard is not compromised by inadvertent ribonuclease contamination. These ARMORED RNA.TM. standards are ideal as RNA standards for the quantification of RNA viruses such as HIV and HCV from human body fluids such as blood and cerebrospinal fluid.

Abstract: The present invention addresses compositions and methods for cleaving nucleic acids. The invention allows one to reliably detecting point mutations in long and short target regions of nucleic acids in a safe, non-labor intensive, and cost effective manner. The methods and compositions of the present invention allow for the use of various RNases, such as RNase A, RNase I and RNase T2, in the detection of mutations. The present invention also identifies reaction conditions that result in significant improvement in specific mismatch cleavage in the NIRCA.TM. assay.

Abstract: The present invention is directed to the process of creating a recombinant nucleic acid standard which is resistant to ribonuclease digestion and is non-infectious. A single strand of recombinant nucleic acid is encapsidated by bacteriophage proteins. The recombinant nucleic acid is a hybrid sequence encoding bacteriophage proteins and a specific non-bacteriophage sequence. A non-bacteriophage RNA sequence can be used as an RNA standard to help quantify the number of RNA molecules in an unknown sample. The recombinant RNA in its packaged form is highly resistant to ribonucleases, insuring that the RNA standard is not compromised by inadvertent ribonuclease contamination. These "ARMORED RNA" standards are ideal as RNA standards for the quantification of RNA viruses such as HIV and HCV from human body fluids such as blood and cerebrospinal fluid.

Abstract: The present invention discloses improved compositions and methods for detecting mutations, including single base changes, in nucleic acid sequences using RNase protection assays. The improvements include concomitant, dramatic reductions in the salt and RNase enzyme concentrations in the RNase digestion reaction mixture which result in greater sensitivity in detecting genetic mutations. Another embodiment of the present invention is kits to be used for the detection of single base mismatches in nucleic acid samples.

Abstract: Disclosed are novel DNA segments, vectors and plasmids containing multiple promoters for use with various polymerases in order to transcribe cloned DNA into RNA. A preferred vector, termed pTRIPLEscript.TM., is described which contains the SP6, T7, and T3 phage promoters in the same orientation and on the same side of a multiple cloning site. This vector efficiently synthesizes in vitro transcripts from all three promoters under conditions of both limiting and saturating nucleotide concentrations. This vector also promotes transcription without crosstalk, i.e., without nonspecific initiation at inappropriate promoters.

Abstract: A method for the recovery of nucleic acids from a reaction mixture, such as nuclease protection assays, comprising the use of one reagent containing a) nuclease for digesting single-stranded RNA and b) a nucleic acid precipitating carrier agent, (sheared DNA or linear acrylamide). A second reagent contains a chaotropic agent (a guanidinium salt) for inactivating said nucleases and an alcohol (ethanol or isopropanol) for the simultaneous inactivation of the nucleases and the precipitation of the nucleic acids without the need for protease digestion or organic extraction.

Abstract: The present invention relates to compositions and methods for increasing the yields of polynucleotide synthetic reactions. In particular, it relates to an improved reaction mixture for use in in vitro RNA transcription and in various other enzymatic reactions in which a polynucleotide is synthesized. The reaction mixture uses high concentrations of total nucleotides, in the order of 12 mM to 40 mM, i.e. levels that were previously thought to be inhibitory. Other useful modifications are also disclosed.