Product Description
Human Survival Motor Neuron 1 (SMN1) Gene Detection Kit (PCR-Melting Curve Method)
General Product Info.:
| Product name |
Technology |
Spec |
Cat.No. |
Cert. |
| Human Survival Motor Neuron 1 (SMN1) Gene Detection Kit |
PCR-Melting Curve Method |
32 |
P124H |
CE |
Spec:
32T/Kit
Intended Use:
This product uses human peripheral EDTA anticoagulant blood as the test specimen to detect the copy number of SMN1 at the 7th exon and (or) the 8th exon in human genomic DNA. Spinal muscular atrophy (SMA) is an autosomal recessive genetic disease characterized by degeneration of motor neurons in the anterior horn of the spinal cord, which is clinically manifested as progressive and symmetrical muscle weakness, atrophy and paralysis in the proximal extremity and trunk. The muscle atrophy is symmetrical, the lower limbs are more serious than the upper limbs, and the proximal part of the body is more susceptible to invasion. Due to the gradual degeneration of muscles, the patient will gradually lose normal physiological functions such as standing, crawling, swallowing, and even breathing, and severe cases will lead to death. The age of onset of this disease is possible from infant to adult, its incidence rate is 1/10000~1/6000, and the population carrying rate is 1/60~1/40.
The main pathogenic gene of SMA is the survival motor neuron (SMN) gene located on chromosome 5q11.2-q13.3. Loss of SMN protein can lead to a large number of early embryonic cell apoptosis. Most cells can tolerate low levels of SMN protein, but spinal anterior horn cells cannot tolerate low levels of SMN protein, which leads to SMA. There are two close and highly homologous SMN genes in the human genome, SMN1 on the telomere side and SMN2 on the centromere side, which differ by only 5 bases, located in exon 7 and 8 and intron 6 and 7, respectively. Both SMN1 and SMN2 can be transcribed. SMN1 is the main pathogenic gene of SMA, and SMN2 is a regulatory gene. According to statistics, about 95% of SMA patients showed homozygous deletion of exon 7 and/or 8 of SMN1 gene, and the remaining 5% or so patients showed heterozygous deletion of exon 7 of SMN1 and point mutation compound heterozygous deletion of SMN1. The main goal of SMA carrier detection clinically is the detection of SMN1 single copy population. This kit detects the copy number of the SMN1 in exon 7 and/or exon 8 of human genomic DNA, and is suitable for screening SMN1 gene carriers in spinal muscular atrophy.
Principle of Test :
Based on real-time fluorescent quantitative PCR, which combines competitive PCR, saturated fluorescent dyes, melting curve analysis technology and normalization software processing technology, this kit is used to detect the copy number of SMN1 in DNA samples. Real-time PCR technology is used to make the target gene and the internal reference gene reach a plateau in the competitive amplification, and the concentration of the amplified product reflects the relative relationship between the copy numbers of the two genes in the substrate template. Saturated fluorescent dyes specifically bind to double-stranded DNA to emit fluorescence, while single-stranded DNA does not produce fluorescence. After melting analysis, through real-time detection of changes in fluorescence signal value during the melting of double-stranded DNA, according to the difference in melting temperature and concentration of different amplified products, the target gene and the internal reference gene form different melting peaks, and the data is normalized by software, P value and R value are calculated, and the copy number of SMN1 is judged based on this. At the same time, negative quality control and positive quality control are set to prevent the occurrence of false negatives and false positives.
Storage Condition & Validity:
All reagents should be stored at -20ºC±5ºCin the dark. The validity period of the reagents is 10 months. The kit should not be frozen and thawed more than 6 times. The opened reagents should be placed at room temperature under the condition of no more than 8 hours. This product is transported under the condition of 0~8ºC. Simulated transport tests indicate that the stability and validity could not be influenced by transport, but the transportation time should not exceed 7 days. Production date and expiration date are shown on the packaging label.
Applicable Instrument:
Gentier 96E real time PCR system from Xi'an Tianlong Science and Technology Co. Ltd., Roche Light Cycler 480 real-time fluorescence quantitative PCR instrument or similar apparatus .
Sample Requirements:
1. Sample: whole blood sample.
2. Collection: 2~5mL of venous blood is extracted from the subject with a disposable sterile syringe, and injected into a glass tube containing EDTA
anticoagulant. The glass tube is immediately reversed gently and mixed for 5~10 times, so that the anticoagulant and venous blood are fully
mixed.
3. It is recommended to use a commercial kit to extract human genomic DNA. The extracted DNA needs to be measured with a UV
spectrophotometer. When 10ng/μL≤DNA concentration requirement≤120ng/μL, OD260/OD280 is within 1.7~2.0. It is recommended that the
extracted DNA be tested immediately, otherwise, please store it below -20°C for no more than 6 months.
Product Leaflet
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