Transferring four 96-well plates to a 384-well plate
This protocol describes the method used to transfer four 96-well plates to a 384-well plate. By using a 384-well plate, more runs can be placed onto the 3700 DNA Analyzer at one time.
- (Note: Due to the limited size of the water and waste reservoir in the 3700 DNA Analyzer, no more than three 384-well plates may be run at any one time.)
- Multimek 96 Automated 96-channel Pipettor (Beckman Instruments)
- Jouan Centrifuge (CR422)
- Rigid Thermo-fast plates, 96 wells (Island Scientific 15-600A) (referred to as 96-well plates)
- 384-well reaction plates (PE Biosystem 4305505)
- Deionized water
- Dried precipitated sequenced PCR products in 96-well plates
- Thermowell sealer (aluminum) for 96-well plates (07-200-684)
- Add 10 l of deionized water to four of the 96-well plates that contain the dried precipitated sequenced PCR products (see Ethanol Precipitation Protocol) and then let this sit at room temperature for 30 min to begin to re-dissolve the sequenced sample.
- Spin the plates down at 1000 rpm (190g) for 20 sec before letting sit for 30 min to re-dissolve.
- After 30 min have elapsed, use the Multimek robot to mix the first 10 l sample. In order to resolve completely, mix sample 30 times using the program "Transfer precipitated product to 384 wells plate."
- After mixing, the robot will transfer the samples from the first 96-well plate into the first quadrant of the 384-well plate
- Repeat steps 3-4 for the next three 96-well plates.
- After the fourth 96-well plate has been added to the 384-well plate, spin the 384-well plate down at 1000rpm (190g) for 20 sec.
- Place a thermowell sealer on top of the 384-well plate and put the 384-well plate onto the loading tray. Make sure that coordinate A1 is on the lower left corner of the loading tray.