KX Genome Walking Kit
- Company Name:Beijing Zoman Biotechnology Co,.Ltd.
- Membership:Free Member
- Registration Date:2018. 07.11
- Contact:Ivory Zhang
- Related Keywords:KX Genome Walking Kit, Genome Walking
Beijing Zoman Biotechnology Co,.Ltd.
Genome Walking is an important molecular biology research technique. It can effectively acquire unknown sequences adjacent to known sequences. Chromosome walking technology mainly has the following applications:
1. According to known gene or molecular markers continuous walking, access to human, animal and plant important regulatory genes, that can be used to study the regulation of structural gene expression. For example, isolate the cloned promoter and study its function.
2. Step-by-step access to non-conserved regions of genes for new species to obtain complete gene sequences.
3. Identification of insertion sites for T-DNA or transposon, identification of insertion sites for foreign genes resulting from transgene technology such as gene gun and transgenesis, etc.
4. Used for gap filling in chromosome sequencing work to obtain a complete genome sequence.
5. Used for overlapping fragmentation of artificial chromosomes PAC, YAC, and BAC.
For a few species (eg, humans, mice, nematodes, rice, Arabidopsis, etc.) that have already been sequenced in the genome, the flanking sequences of a known sequence species can be easily found in the database. However, for most organisms, before knowing their genomic sequence, to know the DNA sequence on both sides of a known region, only chromosome walking technology can be used.
The main problem of chromosome walking based on PCR technology is how to design two specific primers to amplify unknown regions without knowing the unknown region sequence information in advance. Traditional chromosome walking methods, such as reverse PCR and linker methods, have the disadvantages of complicated operation, non-specific amplification, and low connection efficiency.
This kit is efficiently acquiring flanking unknown sequences based on known genomic DNA sequences. Compared to other traditional methods, the kit is characterized by high efficiency, simplicity, high specificity, high sensitivity, and long-term unknown sequence. The main principle is to design three unique primers (SP Primer) with the same direction annealing temperature and higher annealing temperature according to the known DNA sequence, and the four uniquely designed annealing primers with lower annealing temperature provided by the kit（ZFP1, ZFP2, ZFP3, ZFP4, ZFP5, ZFP6, ZFP7, ZFP8, ZFP9）perform asymmetric PCR reactions. In general, randomly selected four kinds of at least one degenerate primer can be used to perform thermal asymmetric PCR reactions with the difference in annealing temperature between the specific primers. The flanking sequences of known sequences can be obtained through three nested PCR reactions. If the length obtained by one experiment does not meet the requirements of the experiment, it is also possible to continue the acquisition of the flanking sequence based on the sequence information acquired by the first walk. In addition, this kit also contains Control DNA and Control Primer for convenient Control experiments.
★ With 9 pairs of random primers, one-time access to the unknown sequence, the results of the authenticity up to 99.99%.
★ High Amplification: The success rate of amplification, product specificity, and output all improved significantly.
★ Amplification of long fragments: The longest step can reach 3.2kb. Usually, it can move about 1.5kb at a time.
★ Improve work efficiency: Clever primer design (without cumbersome experimental steps such as enzyme digestion and adapters), and the entire process of PCR amplification to transformed clones can be completed in 12 hours.
Catalog: ZT601-1; Preps: 10T; Store at -20℃
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