Preparation of 384-Well Plates for Sequencer Loading
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 timeDue 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 Ál 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 sample down at 1000 rpm (190g) for 20 sec before letting sit for 30 min in freezer.
- After 30 min have elapsed, use the Multimek robot to mix the 10 Ál sample 30 times in order to resolve completely.
- After mixing, the robot will transfer the samples from the first 96-well plate into the first quadrant of the 384-well plate
- Repeat step 2-3 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.