The present disclosure provides chimeric primers suitable for use in the amplification of a nucleic acid sequence. In some aspects, these chimeric primers reduce the formation of primer dimers and/or off-target amplification products, compared to amplification reactions carried out using unmodified primers.

The present disclosure relates to the field of biotechnology, in particular, to a combination product for detecting DNA. The product includes ribonuclease H II and a probe; the probe is a single-stranded probe and has a sequence which can be partially or entirely complementary to a target DNA molecule to be detected, and one or more RNA bases are embedded in the complementary region of the probe and divide the complementary region of the probe into at least two segments, each of which independently has DNA bases and base substitutions of less than or equal to 13 in total.

The present disclosure relates to the field of biotechnology, in particular, to a combination product for detecting DNA. The product includes ribonuclease H II and a probe; the probe is a single-stranded probe and has a sequence which can be partially or entirely complementary to a target DNA molecule to be detected, and one or more RNA bases are embedded in the complementary region of the probe and divide the complementary region of the probe into at least two segments, each of which independently has DNA bases and base substitutions of less than or equal to 13 in total.

A method for specifically and efficiently quantifying the expression of targeted RNA variants with specific terminal sequences suitable to identify multiple isoforms bearing complex heterogeneity in terminal sequences by hybridizing a 5′-Dbs-adapter to the 5′-end of target RNAs, wherein the 5′-Dbs-adapter has a stem-loop structure whose protruding 5′-end base-pairs with the 5′-end of target RNAs, and wherein the loop region of 5′-Dbs-adapter contains a base-lacking spacer which will terminate reverse transcription in a subsequent step; hybridizing a 3′Db-adapter to the 3′-end of target RNAs, wherein the 3′-Db-adapter has a stem-loop structure whose protruding 3′-end base-pairs with the 3′-end of target RNAs; ligating both adapters with target RNAs by RN12 ligation to form a “dumbbell-like” structure; and, amplifying and quantifying the ligation product by RT-PCR.

Provided are methods for preparation and analysis of nucleic acids. Some embodiments include reverse transcribing the RNA with barcoded primers to produce cDNA while maintaining the DNA in the sample, sequencing the DNA and cDNA together, and differentiating the sequenced DNA and cDNA using the barcode or barcodes of the primers. Some embodiments include analyzing the DNA and cDNA sequences of multiple samples separating reads into k-mers, and comparing the k-mers between samples to identify differential sequences between the sequences of the samples.

Provided are methods for preparation and analysis of nucleic acids. Some embodiments include reverse transcribing the RNA with barcoded primers to produce cDNA while maintaining the DNA in the sample, sequencing the DNA and cDNA together, and differentiating the sequenced DNA and cDNA using the barcode or barcodes of the primers. Some embodiments include analyzing the DNA and cDNA sequences of multiple samples separating reads into k-mers, and comparing the k-mers between samples to identify differential sequences between the sequences of the samples.

The present disclosure relates, in part, to improved methods of making single-stranded DNA (ssDNA) from double-stranded DNA (dsDNA), as well as use of the resulting ssDNA for genome engineering. The disclosure also relates, in part, to improved methods of genetic modification using single stranded DNA binding proteins.

The present disclosure relates, in part, to improved methods of making single-stranded DNA (ssDNA) from double-stranded DNA (dsDNA), as well as use of the resulting ssDNA for genome engineering. The disclosure also relates, in part, to improved methods of genetic modification using single stranded DNA binding proteins.

The present invention may provide a small RNA detection sensor comprising: at one end thereof, a first sensing region comprising nucleotides having a sequence complementary to target small RNA; and a PCR-capable region that is coupled to the first sensing region, the small RNA detection sensor to synthesize a replication region complementary to the PCR-capable region by a DNA polymerase by using the target small RNA as a primer, and amplify the PCR-capable region and the replication region.

The present invention relates to a method for preparing a DNA oligomer into which a single nucleotide is incorporated using a terminal deoxynucleotidyl transferase. According to the present invention, by using a base hydrolysis reaction or a ribose-borate complex formation method, single incorporation of normal and modified nucleotides in a TdT enzyme-based DNA oligomer modification method can be facilitated. In addition, the method simultaneously provides the usability of TdT and the quantitativeness of modification group incorporation, thereby being effectively usable in the development of a quantitative detection technique or in a crosslinking reaction, both of which require quantitativeness and, according to the present invention, a DNA oligomer, into which a single nucleotide which is a product of a TdT reaction is incorporated, has an exposed a 3′ hydroxyl terminus, which is an enzyme recognition site, such that an additional enzymatic technique such as primer extension or ligation can be introduced.