How to Set up a Sanger Sequencing Run - Seq It Out #16
Protocol
Difficulty
intermediate
Steps
1
Select the capillary array length
Choose the appropriate capillary array based on your application. For standard sequencing runs, select the 50 cm array, while 36 cm arrays are used for fragment analysis or human identification applications.
▶ 00:26
2
Install the polymer pouch
Select the correct polymer based on your application needs. For this standard run, use POP-7 polymer and snap it into the instrument frame.
▶ 01:20
3
Prepare cathode and anode buffers
Remove the seals from the pre-diluted buffer containers and add septum to the cathode container. Place the cathode buffer in the designated cathode position and the anode buffer in the anode position.
▶ 01:54
4
Verify RFID tracking information
Confirm that all consumables are RFID-labeled to enable automatic tracking of part numbers, lot numbers, expiration dates, and usage information throughout the run.
▶ 02:29
5
Launch data collection software
Start the Data Collection software on the 3500 Genetic Analyzer, ensuring you are using the latest version available.
▶ 02:52
6
Fill array and perform calibration
Fill the capillary array with polymer and complete a spatial calibration to prepare the instrument for sequencing.