• TB Typing Kit(VNTR-9)
  • TB Typing Kit(VNTR-9)

TB Typing Kit(VNTR-9)

This kit is based on the latest research progress in molecular epidemiology and has been optimized to form a genotyping product for Mycobacterium tuberculosis. This product uses the variable number tandem repeats (VNTR) polymorphism in the genome of Mycobacterium tuberculosis to genotype and differentiate clinical strains
$8.50
  • TB Typing Kit(VNTR-9)

SPECIFICATION



                        TB Typing Kit(VNTR-9)

This kit is based on the latest research progress in molecular epidemiology and has been optimized to form a genotyping product for Mycobacterium tuberculosis. This product uses the variable number tandem repeats (VNTR) polymorphism in the genome of Mycobacterium tuberculosis to genotype and differentiate clinical strains, which is a powerful tool for studying the molecular epidemiology of Mycobacterium tuberculosis and monitoring the transmission of tuberculosis. Compared with other existing VNTR based typing systems for Mycobacterium tuberculosis, this typing system has stronger discrimination ability for strains prevalent in China, making it particularly suitable for the needs of Chinese users.
By carefully optimizing the primer sequences of each PCR reaction and the composition of the premixed reaction solution, this kit has strong anti-interference ability. Compared with non user prepared reagents, this product significantly improves the strength of specific band signals, reduces the occurrence rate of non-specific bands when using crude templates (boiled bacterial solution), makes experimental operations more convenient and efficient, and improves the detection success rate. The premixed reaction solution of this product has good chemical stability and can effectively resist repeated freeze-thaw cycles (10 times) and prolonged room temperature environments (one week), better adapting to the flexibility needs of users in testing work.
This kit integrates the identification of Mycobacterium tuberculosis (MTBC), non Mycobacterium tuberculosis (MOTT) and template quality, identification of Mycobacterium tuberculosis Beijing family strains, and subsequent 9-site VNTR typing into one kit. Users only need one kit and 12 PCR reactions to obtain the complete information required for genotype identification of the tested strain. The Hunter Gaston index (HGI) for detection can reach 0.989. Moreover, the results of genotype identification can be digitized and recorded, making it easy for experimental data from different batches of samples, regions, time periods, and researchers to be compared and analyzed, greatly improving the efficiency of data utilization.
Application of component specifications
Mtb identification of 1mL to determine whether the strain belongs to Mycobacterium tuberculosis human type
Quality Identification of 16S rRNA 1mL DNA Template
RD105 1mL identifies whether the strain belongs to the Beijing genotype
QUB-11b 1mL 9-site VNTR genotyping detection
QUB-18 1mL
QUB-26 1mL
MIRU26 1mL
MIRU31 1mL
MIRU40 1mL
Mtub21 1mL
Mtub04 1mL
VNTR2372 1mL
Marker I 1mL DNA Molecular Weight Standard I
Marker II 400 μ L DNA Molecular Weight Standard II
Storage: -20 ℃, valid for one year.
To avoid contamination, it is recommended to prepare samples and prepare PCR Mix in different locations and use different pipettes.
Attention should be paid to labeling in all stages of sample DNA collection, extraction, and amplification, while preventing cross contamination between different samples.
Commonly used reagents and consumables need to be high-pressure sterilized before the experiment.
Each PCR Mix tube contains different primers and cannot be mixed. It can be divided into different quantities at once according to experimental needs to avoid repeated freeze-thaw.
To avoid splashing of the reaction liquid when opening the reaction tube, please briefly centrifuge before opening the cover and collect the liquid at the bottom of the tube. If it accidentally splashes onto gloves or tabletop, immediately replace the gloves and wipe the tabletop with 75% alcohol or dilute acid.
When aspirating, be careful not to cross contaminate PCR Mix. It is recommended to wipe the pipette head twice with 75% alcohol before taking Mix each time.
Preparation before the experiment: 1 × TE buffer (pH=8.0), 0.5 × TBE buffer, agarose, ethidium bromide (EB), ordinary PCR instrument, DNA electrophoresis equipment and gel imager, 0.2ml PCR reaction tube, eight array or 96 hole PCR tube, pipettes of different specifications: 0.5~10 μ L and 20-200 μ L.
1、 DNA template preparation:
Scrape a small amount of (1-2 inoculum rings) sample from the solid culture medium and resuspend on 100 μ In LTE, inactivate at 80 ℃ for 30 minutes.
The inactivated strains were taken out of the P3 laboratory for the following operations:
Boil at 100 ℃ for 10 minutes (the cover of the EP tube may burst during boiling, avoid it as much as possible, fasten the EP tube, and do not let water enter the tube), immediately place it on ice for 2 minutes at 12000 rpm (~13400 × g) After centrifugation for 10 minutes, take the supernatant and place it in another sterile EP tube, mark it, and store it at -20 ℃.
2、 Detection program:
Take out the TB genotyping kit VNTR-9, wait for the liquid to equilibrate to room temperature, gently shake 3-4 times, mix well, centrifuge at 12000 rpm for 5 seconds, and collect the liquid covered into the tube.
Genotype analysis of Mycobacterium tuberculosis:
Identify whether the sample is human type Mycobacterium tuberculosis (important! This step cannot be omitted!):
PCR amplification: reaction system is 20 μ L.
Add 19 to each PCR tube separately μ L M
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