Views: 10 Author: Site Editor Publish Time: 2022-01-11 Origin: Site
PCR Identification Kit Features:
1. One-tube operation, users only need to provide samples.
2. Primers are designed according to the conserved region of soybean heat shock protein gene, which can specifically detect soybean components in the sample, but cannot detect other non-soy components.
3. Fast, the entire detection process (according to one sample) takes only 2.0 hours.
4. Only need common PCR machine and gel electrophoresis machine, no need to configure expensive equipment.
5. The lower limit of detection for soybean components in mixed samples is 0.1%, and the lower detection limit for nucleic acids for soybean components in samples is 1.0ng/μL.
6. This product can only be used for scientific research, enough for 50 times of routine PCR in 40uL system.
How to use PCR identification kit:
1. Dilute the standard curve samples (take 10E2-10E7 copies/μL as an example of six 10-fold dilutions). Since the standard concentration is very high, the following dilution operations must be performed in a separate area, and must not contaminate the sample or other components of the kit).
1.1 Label the 6 centrifuge tubes as 7, 6, 5, 4, 3, and 2.
2.1 Use a cored pipette tip to add 45 μL of fluorescent PCR template dilution solution, *use a cored pipette tip, the same below).
3.1 Add 5 μL of positive control (concentration of 1×10E8 copies/μL, provided by the kit) to the No. 7 tube, shake well for 1 minute, and obtain a standard curve sample of 1×10E7 copies/μL. Set aside on ice.
4. 1Change the pipette tip, add 5 μL of 1×10E7 copies/μL positive control (obtained in the previous step) to tube 6, shake well for 1 minute, and obtain a standard curve sample of 1×10E6 copies/μL. Set aside on ice.
5.1 Change the pipette tip, add 5 μL of 1×10E6 copy/μL positive control (dilution in the previous step) to tube 5, and shake well for 1 minute
clock to obtain a standard curve sample of 1×10E5 copies/μL. Set aside on ice.
6. 1Repeat the above procedure until 6 dilutions of the standard curve samples are obtained. Set aside on ice.
2. Preparation of sample DNA
2.1 Purify the DNA of the sample by the method of your choice. This product is compatible with most nucleic acid purification products on the market.
2.2 If there are N samples, N+2 sample extractions need to be performed, one extra for the sample preparation positive control tube and the other for the sample preparation negative control tube.
2.3.Set up the qPCR reaction (20μL system, in the sample preparation room)
2.4. If doing quantitative analysis and only doing 1 replicate, label N+9 PCR tubes, of which N+2 are for the N+2 samples from the previous step and 1 is for the PCR negative control (use water as template). ), 6 for the standard curve. If qualitative analysis is performed and only 1 repetition is performed, label N+4 PCR tubes, of which N+2 are used for the N+2 samples obtained in the previous step, and 1 is used for PCR negative control (use water as template) , 1 for PCR positive control.