Transgenic maize line TC1507 nucleic acid detection kit instruction manual

Transgenic Maize Line TC1507 Nucleic Acid Detection Kit (PCR-Fluorescence Probe Method)

â—† Product Description

The transgenic detection series can amplify specific nucleic acid fragments of transgenic components in food, feed and other samples, and judge the results by real-time amplification curve. This product is used for the detection of transgenic maize line TC1507 with a detection limit of 0.1% .

â—† Product composition (48 test)

â—† Applicable instruments

A PCR instrument such as a fluorescent PCR machine (ABI 7500, CFX 96, Mx 3005P, LineGene 9600).

â—† Self-supplied supplies and instruments

1 sterilize 1.5mL or 2.0mL centrifuge tube; 2 sterilize 0.2mL PCR tube or octa tube; 3 ice box; 4 pipette (11-10μL, 10-100μL, 100-1000μL) and matching sterilization tips 5 centrifuge; 6 vortex mixer; 7 metal bath; 8 homogenizer, mixer or mortar and other grinding equipment; 9 electronic balance.

â—† Notes

1. This reagent has high detection sensitivity. In order to prevent pollution, the experiment is to be partitioned.

1) First zone: reagent preparation zone.

2) Second zone: sample preparation area.

3) Zone 3: Amplification and product analysis zone.

★ It is best to physically isolate the partitions to avoid contamination caused by human factors.

2. Wear work clothes and latex gloves during the experiment, use pipettes, and treat all waste generated in the test in time.

3. Strictly follow the operation steps, reagent preparation and sample loading steps should be carried out on ice in strict accordance with the instructions.

4. The components in the reaction solution are sensitive to light and should be stored away from light. The reagent should be completely thawed before use, but repeated freezing and thawing should be avoided. It is recommended to centrifuge for 30 seconds before use, and store the reaction solution in an appropriate volume according to the frequency of detection.

5. After the reaction is completed, the expansion tube should be placed in a sealed bag and discarded. On the same day, the lid is opened and the aerosol is easily contaminated. It is forbidden to open the lid.

6. Do not mix and use different batches of reagents and use them within the validity period.

â—† Sample processing

Samples prepared by the method of GB/T 19495.3-2004 Detection of nucleic acid extraction and purification of genetically modified products or other standards are prepared and used for storage.

For detailed steps, please follow the standard operation or check the food safety software.

â—† Experimental operation

The reagents were completely thawed and the components were centrifuged for 30 s.

1. Reagent preparation (reagent preparation area, placed in an ice box):

If there are N samples to be tested, refer to the table below, calculate the amount of reaction solution according to N+2 (N samples to be tested + 1 negative control + 1 positive control), vortex and mix, centrifuge for 30 s, Dispense in 0.2 mL PCR tubes.

2. Template preparation (sample preparation area)

It is recommended to use the supporting plant genomic DNA extraction series products. The specific process is detailed in the product manual.

3. Add a template (sample preparation area, placed in an ice box)

5 μL of the template was added to the PCR tube containing the reaction solution in the first step, and the order was NG-P, the sample template to be tested, and PG-1507-P. The mixture was vortexed for 30 s, centrifuged for 1 min, and the PCR amplification reaction was immediately performed.

4. Amplification reaction (amplification and product analysis area)

Using a real-time PCR instrument, the fluorophore was selected for FAM and the quencher group was selected for TAMRA.

Set up the amplification reaction according to the following conditions:

5. Baseline and threshold settings

The baseline adjustment takes 3-15 cycles of fluorescence signal, and the threshold line should exceed the highest point of the negative control amplification curve.

â—† Result judgment

The test sample has no Ct value, the curve is straight or slightly oblique, there is no "S" type amplification curve, the sample can be reported negative, does not contain the transgenic maize line TC1507 or the content is below the detection limit;

The test sample Ct ≤ 40, the curve is an "S" type amplification curve, which can directly report the sample positive, containing the transgenic maize line TC1507;

Test sample 40

• The NG reaction is a smooth straight line, and the PG reaction is an "S" type amplification curve. The test result is valid, otherwise it is invalid. If the duplicate test results are still invalid, please contact technical support.
• 

Peptides Products

An organic compound formed by dehydration of amino acids, containing carboxyl and amino groups, and being amphoteric. Also known as "peptide".
Two, one of the amide. It consists of two or more amino acids joined by the amino group of one amino acid to the carboxyl group of the other. An amino acid cannot be called a peptide, nor can a peptide be synthesized, but must be a compound of two or more amino acids linked by a peptide bond. Compounds where two amino acids are linked by a peptide bond are called dipeptides; A compound with three amino acids linked by peptide bonds is called a tripeptide, and so on, a compound with thirty-four amino acids linked by peptide bonds is called a thirty-four peptide.
A peptide is a chain of polymers of amino acids
A peptide is a chain of polymers of amino acids

Three, is related to the biological body of a variety of cell functions of biological active substances. Hundreds of peptides have been found in living organisms, which are essential participants in the body's complex physiological activities. All cells can synthesize peptides and their functional activities are regulated by peptides. It involves hormones, nerves, cell growth and reproductive fields, and its importance lies in the regulation of various systems, organs and cells in the body. The physiological and pharmacological effects of enzymatic polypeptides are mainly to activate related enzymes in vivo, promote the permeability of intermediate metabolic membrane, or affect specific protein synthesis by controlling DNA transcription or translation, and eventually produce specific physiological effects or play their pharmacological effects. Peptide is better than amino acid, one is faster than amino acid absorption; Second, it is absorbed by the body in a complete form; Third, active absorption (amino acids are passive absorption); Fourth, low consumption, compared with amino acids, peptide absorption has the characteristics of low consumption or no energy consumption, peptide absorption through the duodenum, directly into the blood circulation, their own energy nutrition transported to various parts of the human body; Fifth, peptide absorption is less than amino acid, with the characteristics of unsaturated; Sixth, there are only 20 kinds of amino acids, with countable functions, while peptides take amino acids as substrates, which can be synthesized into hundreds of thousands of kinds.