Sequencing and Chip Technology_Custom Services

Location: Home > Custom Services > Sequencing and Chip Technology

PCR-seq

Introduction Specific DNA segments defined by the sequence of two oligonucleotides can be enzymatically amplified up to a millionfold using the polymerase chain reaction (PCR). One of the most significant uses of this technique is for the generation of sequencing templates, either from cloned inserts or directly from genomic DNA. To avoid the problem of reassociation of the linear DNA strands in the sequencing reaction, ssDNA templates can be produced directly in the PCR or generated directly from dsDNA by enzymatic treatment, electrophoretic separation or affinity purification. By combining PCR with direct sequencing, both the amplification and the sequencing reaction can be performed in the same vial. Finally, the use of fluorescently labeled terminators or sequencing primers will allow the whole procedure to be amenable to complete automation. One of the main advantages of sequencing a PCR product directly is that you will not see PCR-generated mutations. The reason for this is that you have a large population of templates in your initial PCR reaction and even a mutation introduced in one PCR product in the first round of amplification will only be prese...

Details >>
BAC Library Sequencing

Introduction Bacterial Artificial Chromosome (BAC) is a DNA construct, based on a functional fertility plasmid (F-plasmid), used for transforming and cloning in bacteria, usually E. coli. BACs are often used to sequence the genome of organisms. A short piece of the organism's DNA is amplified as an insert in BACs, and then sequenced. Finally, the sequenced parts are rearranged in silico, resulting in the genomic sequence of the organism. As a Large-insert genomic DNA libraries in bacteria, BAC libraries could provide a way to divide the complexity of the human genome into a composite of large DNA segments of reduced complexity. Ideally, BAC libraries should completely represent the genome without cloning artifacts or rearrangements and should be provided in an addressable format with clones physically separated. Today, BAC libraries are used as a source of substrates for shotgun sequencing projects to create a database of end sequences and restriction fingerprints for building overlapping clone sets, they also can provide scaffolding information for mapping sequence contigs to localized genomic regions by using a direct genomic shotgun sequencing appr...

Details >>
Plasmids DNA Sequencing

Introduction DNA sequencing provides the most complete characterization of recombinant plasmid DNAs. Using primers targeting the plasmid backbone or the insert sequence, the identity and order of nucleotide bases for any given DNA can be determined. In the context of cloning, sequencing allows users to confirm the DNA sequence of the insert, insert orientation, and to examine the junctions between the plasmid and insert DNA. Sanger sequencing technique is the most commonly used method for determining DNA sequences of recombinant plasmids. Sanger sequencing involves the use of a DNA polymerase, a primer, unlabeled deoxynucleotide triphosphates (dNTPs), and fluorescently labeled dideoxynucleotide triphosphates (ddNTPs), where each base is labeled with a unique fluorophore. Incorporation of a ddNTP into the newly synthesized strand prevents the addition of subsequent nucleotides, stopping the further elongation of the DNA molecule and resulting in a DNA product with a fluorescently labeled ddNTP at the end of the strand. The sequence of nucleotides can then be determined by separating the DNA products by size.

Details >>
Sanger Sequencing

Introduction Sanger sequencing is the process of selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase during in vitro DNA replication; it is the most widely used method for the detection of SNVs. Sanger sequencing is the most widely used sequencing method for smaller-scale projects, validation of NGS results and for obtaining especially long contiguous DNA sequence reads (> 500 nucleotides).

Details >>
Real Time PCR

Introduction Quantitative Real-Time PCR (qRT PCR) technology refers to the method of instantaneously measuring the amount of specific products by continuously monitoring the change of fluorescence signal strength during PCR exponential amplification, and finally quantitatively analyzing the unknown template by standard curve. Qualitative analysis: Test the sample for a particular gene; Absolute quantification: Test the sample for a particular gene and determine the copy number of the gene in the sample; Relative quantification: Test the sample for a particular gene and determine the relative expression of the gene compared to a certain housekeeping gene and the control group; ☀ Customers should offer: Samples: Please provide fresh and sufficient samples, tissue >100~200 mg, RNA or DNA template >5 μg; Other information: Please provide objective gene sequence, name and gene accession number, etc.; primer sequences or primers; detailed background information; sample source and its abundance, related literature, etc.; ☀ Deliver the finished products: ...

Details >>
Chemical synthesis of miRNA

Introduction In recent years, artificially synthesized miRNAs (amiRNAs) have been successfully applied to the expression of silent target genes and their functional studies. And they can specifically silence a single gene, and simultaneously silence multiple related but different genes. miRNA products mainly include miRNA mimics and miRNA inhibitors. First of all, miRNA mimics can mimic the high level expression of endogenous mature miRNAs in cells, and further enhance the silencing of them. Besides, it can also reduce the amount of intracellular protein expression, and be used to perform gain-of-function studies. What’s more, miRNA inhibitors can specifically target and knock out single miRNA molecule, attenuate the gene silencing effect of endogenous miRNAs, and thus it increases protein expression, and helps to carry out loss-of-function studies, in which researchers can screen miRNA target sites, and screen of miRNAs that regulate the expression of a gene and the ones affecting cell development. Service name Specifications （OD） ...

Details >>

Contact Us

Inquired items

Inquired items

All Departments