The present disclosure provides methods and kits for performing high multiplex PCR using molecular barcodes. The methods disclosed herein separately extend a set of primers (BC primers) that each comprise a target-specific sequence, a molecular barcode and a universal sequence, and amplify the resulting extension products using another set of primers (LA primers) that each comprise another target-specific sequence and a universal sequence. The methods may further comprise amplification using universal primers (preferably comprising an adapter).

The present disclosure relates to the treatment of Hutchinson-Gilford Progeria Syndrome (HGPS) and diseases related to vascular ageing and in the treatment of smooth muscle cells diseases, in particular an inhibitor of a metalloprotease the treatment of smooth muscle cells diseases. The disclosure subject matter describes a more effective therapies for the treatment of Hutchinson-Gilford Progeria Syndrome and diseases related to vascular ageing, or namely by the use of an inhibitor of a metalloprotease.

Disclosed are methods of measuring the amount of exposure of a host to a DNA double-stranded break (DSB)-causing agent by determining the ratio of the quantity of γ-H2AX to the quantity of total H2AX in a biological sample from the host as compared to the ratio of the quantity of γ-H2AX to the quantity of total H2AX in a positive control biological sample that has been exposed to a known amount of a DSB-causing agent. Related kits and methods of quantifying DSBs in a test biological sample are also disclosed.

A nucleic acid amplification method includes a step of heating a first region of a container housing a droplet containing a target nucleic acid and a sample necessary for amplification of the target nucleic acid to a denaturation temperature of the target nucleic acid and heating a second region different from the first region to a synthesis temperature of the target nucleic acid, and an amplification step of repeating a cycle through a denaturation stage at which the droplet housed in the container is moved to and retained in the first region and a synthesis stage at which the droplet is moved to and retained in the second region at a plurality of times. At the amplification step, periods of part of cycles of the plurality of cycles are made shorter than periods of the other cycles.

The lack of clear predictors of prostate cancer progression leads to subjective decision-making regarding courses of treatment. The identification of new biomarkers that are predictive of recurrence after radical prostatectomy would advance the field of prostate cancer treatment. Disclosed are miRNAs that can be used as molecular biomarkers to detect or predict the progression of prostate cancer and to adjust a treatment plan accordingly. Furthermore, kits are included for the detection of these miRNAs.

The present disclosure encompasses methods of error corrected sequencing (ECS) that enable detection of very rare mutations well below the error rate of convention next generation sequencing (NGS). Further, the methods disclosed herein enable multiplex targeting of genomic DNA.

The present invention aims at providing a specific antibody that can simply and rapidly detect Mycoplasma pneumoniae which is a causative bacterium of mycoplasma pneumonia, with high sensitivity, and also an immunological detection method and a kit containing the same antibody. The present invention makes it possible to diagnose infection with Mycoplasma pneumoniae more rapidly and specifically than the conventional method, by producing an antibody recognizing a specific epitope of P30 protein of Mycoplasma pneumoniae and performing an immunological detection using the antibody. Also, the present invention enables easy and rapid detection of Mycoplasma pneumoniae and diagnosis of infection with the same at a hospital or the like without need of specialized instruments or skilled techniques.

The present invention relates to a primer for PCR obtained by, directly or through inosine as a linker, linking a complementary nucleotide sequence or a complementary nucleotide sequence including a mis-matched nucleotide sequence to the 5′-terminal of a forward or reverse primer; and to a PCR method including a step of mixing a nucleic acid template in a PCR composition including the primer and then performing PCR on the mixture. The primer for PCR of the present invention includes a complementary nucleotide sequence or a mis-matched nucleotide sequence in a complementary nucleotide sequence, which is linked to the 5′-terminal thereof directly or via a linker, thereby lowering the sensitivity increase due to the increase in amplification products and reducing non-specifically occurring reactions in PCR.

The present invention relates to a primer for PCR obtained by, directly or through inosine as a linker, linking a complementary nucleotide sequence or a complementary nucleotide sequence including a mis-matched nucleotide sequence to the 5′-terminal of a forward or reverse primer; and to a PCR method including a step of mixing a nucleic acid template in a PCR composition including the primer and then performing PCR on the mixture. The primer for PCR of the present invention includes a complementary nucleotide sequence or a mis-matched nucleotide sequence in a complementary nucleotide sequence, which is linked to the 5′-terminal thereof directly or via a linker, thereby lowering the sensitivity increase due to the increase in amplification products and reducing non-specifically occurring reactions in PCR.

The present disclosure provides compositions and methods for labeling a single stranded target nucleic acid. Subject compositions include a Cas9 protein, a Cas9 guide RNA, and a quenched PAMmer. A subject quenched PAMmer is a single stranded oligonucleotide having (i) a protospacer adjacent motif (PAM) sequence; (ii) a detectable label; (iii) a quencher moiety that quenches the detectable label; and (iv) at least one of: a specificity segment positioned 5 of the PAM sequence, and an orientation segment positioned 3 of the PAM sequence. In the subject methods, the Cas9 protein cleaves the quenched PAMmer at a cleavage site positioned between the detectable label and the quencher moiety to produce: (a) a first cleavage product that is hybridized with the target nucleic acid and comprises the detectable label; and (b) a second cleavage product that is not hybridized with the target nucleic acid and comprises the quencher moiety.