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DNA sequencing (or translated DNA sequencing) refers to the analysis of the base sequence of specific DNA fragments, that is, the (G) arrangement of adenine (A), thymine (T), cytosine (C) and guanine. The emergence of rapid DNA sequencing methods has greatly promoted the research and discovery of biology and medicine.

DNA sequence knowledge has become an indispensable knowledge in basic biological research and in many application fields, such as diagnosis, biotechnology, forensic biology and biosystematics. The rapid sequencing speed with modern DNA sequencing technology has helped to achieve complete DNA sequencing, or multiple types of genome sequencing and complete DNA sequences of life species, including human genome and many other animal, plant and microbial species.

Generate several groups of radiolabeled oligonucleotides that are independent of each other. Each group of oligonucleotides has a fixed starting point, but they randomly terminate on a specific residue or residues. Since each base on DNA has an equal opportunity to appear at the variable termination end, each group of products above is a mixture of oligonucleotides, and the length of these oligonucleotides is determined by the position of a specific base on the whole segment of the original DNA. Under the condition that different DNA molecules with only one nucleotide difference in length can be distinguished, electrophoretic analysis is carried out for each group of oligonucleotides. As long as several groups of oligonucleotides are added to several adjacent swimming lanes in the sequencing gel, the nucleotide sequence on DNA can be read directly from the radioactive film of the gel.
Routine DNA sequencing Sequencing of complex structures Phage sequencing Primer Walking
For routine sequencing samples of unknown concentration, you can choose this service. The types of samples include plasmids, PCR products (purified PCR or PCR stock solution without heterobands), bacteria (plate cloning or glycerol bacteria), and bacteriophages with circular genomes (supernatant or plaque).

Due to the influence of secondary structure or other internal factors, complex structure is a huge challenge for sequencing. Conventional sequencing schemes are prone to early interruption or disordered sequencing results.

The samples with the following sequence characteristics are suitable for the sequencing scheme for complex structures:

High GC content: templates with GC content>60%, or templates with local high GC content

Hairpin structure: it contains two reverse repeat sequences with at least 3 bases between them

Repetitive sequence: multiple consecutive repeats of 2 or 3 bases.

DNA hairpin structure and RNAi template sequencing

RNA interference (RNAi) is a mechanism that specifically inhibits the expression of a related gene mediated by double stranded RNA (dsRNA). It has gradually developed into a fast and effective emerging technology to study gene function.

RNAi template brings great challenges to sequencing due to its "card issuing structure". Small hairpin RNA (shRNA) used in RNAi research usually cannot be sequenced successfully, and the sequencing results are often interrupted in advance or the signal is disturbed. Qiangyao Bio has a sequencing program specifically for shRNA templates, and the sequencing results have been unanimously recognized by customers.

The sequencing scheme of proboscis can directly sequence phages (phage supernatant or plaque) with circular genome. When it is difficult to obtain enough phage DNA samples for sequencing, this operation can save you the tedious process of DNA preparation and obtain high-quality sequencing results.

This service includes:

·Save time and cost - no need for bacterial culture and DNA preparation

·Sequence analysis by ABI3730xl DNA sequence analyzer

·Phred20 effective length is greater than 600bp

·Provide universal primers for powerful organisms for free

·You can choose to submit your order online. Qiangyao Bio CLIMS system will save your order information and sequencing results for a long time:

·The technical support of Qiangyao Biotech will communicate with you in time for the problems in the project

QYAOBIO can be sequenced by Primer Walking to help you obtain the sequence information of unknown clones or verify the accuracy of cloned sequences. If you provide reference sequences, we can complete the full length sequencing of long fragment clones at one time.

This service includes:

·Primer design and synthesis

·Manual editing according to sequence map

·Sequence splicing

·Send splicing sequence results