ABI 7300 Real Time PCR System Real

ABI 7300 Real-Time PCR System Real-time PCR analysis system

The main features of this device:

1. The quantitative detection: absolute quantification (pathogen detection, detection of genetically modified foods, gene expression studies);

Relative quantitation (differentially expressed genes in different tissues, drug efficacy assessment).

2. Point mutation detection: alleles associated with the disease mutation detection; SNP detection.

ABI 7300 Real-Time PCR System Real-time PCR procedures

First, the instrument on / off procedures:

1. The boot process

1.1 Turn on the computer.

1.2 PCR instrument to open the power switch (Note: The instrument has two power switches, a control PCR, and a source control system).

Double-click on the software icon on the desktop 1.3, click "yes", and enter the software's main menu.

2.shutdown procedures:

2.1 save data and exit the software's main menu.

2.2 Turn off the computer, turn off the power switch PCR instrument.

Second, the setup program and open the file

1. The experimental program has been pre-set

Click Software 1.1 left home directory "Library", after entering the interface.

1.2 Select "View Protocol" page, according to the path, find the stored files.

2. Create a new program file

Click Software 2.1 left home directory "Workshop", enter the interface.

2.2 Select "Edit Protocol" page, enter the appropriate temperature, time, repeated cycles (Repeats), add or remove steps (Cycle and Step), in the "Select data collection step (s)", select the desired fluorescence collection steps.

2.3 When complete, enter the program in the "Protocol Filename" in the file name, click on "Save this protocol".

Third, the instrument operating procedures:

1. Edit the sample :

PCR 1.1 containing the reaction solution into the PCR reaction tube, record the sample placed an order.

1.2 Click on the left into the software home directory "Workshop" screen, select "Edit Plate Setup" page.

1.3 Click "Samples: Whole Plate loading", based on the sample placed the order and sample properties, select the corresponding icon (Unknown and Standard), the corresponding position on the plate for identification is standard (Standard), and at the bottom right of the screen the "Standard Quantity", enter the appropriate concentration.

1.4 Click the "Select and load fluorophores", select the fluorescent marker "1. Or deselect a fluorophore", select the color corresponding to the mark in the "2.Assign a color", and then the holes in the plate selection 1.3 In fluorescent markers.

1.5 After the completion of the "Plate Setup Filename:" Enter the sample file in the file name, click on "Save this plate setup".

2. Run

2.1 After completion of editing samples, in which the page click on "Run with selected protocol".

2.2 Transfer "Run Prep" page, confirm "Select run purpose" is selected "PCR Quantification Melt Curve" and "Select well factor source" is selected "Experimental Plate", check it "Selected Protocol" and "Selected Plate "file name is correct.

2.3 And then enter the "Sample volume" in the reaction system, click on "Begin Run", enter the file name you want to save the data in the dialog box, click Save.

3. Data analysis

Click Software;

3.1 left home directory "Library", enter "View Post-Run Data" page.

3.2 Locate and select the file you want to analyze the data, click on "Analyze Data", into the "Data Analysis" interface.

3.3 According to "baseline" and "Threshold" analysis principles, data analysis, observed PCR Standard

Curve ".