SOLiD™ EZ Bead™ Amplifier

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[править] Materials and equipment required

  Order one or more of the kits according to the scale of the ePCR.

Item (part number) Components
SOLiD™ EZ Bead™ Amplifier E10 Accessories Kit (4453071)
•SOLiD™ EZ Bead™ Amplifier E-Small Pouch Pack
•SOLiD™ EZ Bead™ Amplifier E-Small Funnel Pack
SOLiD™ EZ Bead™ Amplifier E20 Accessories Kit (4453077)
•SOLiD™ EZ Bead™ Amplifier E-Small Pouch Pack
•SOLiD™ EZ Bead™ Amplifier E-Small Funnel Pack
SOLiD™ EZ Bead™ Amplifier E80 Accessories Kit (4453072)
•SOLiD™ EZ Bead™ Amplifier E-Small Pouch Pack
•SOLiD™ EZ Bead™ Amplifier E-Small Funnel Pack
SOLiD™ EZ Bead™ E10 System Consumables, 4-pack (4453093)*
•SOLiD™ EZ Bead™ Emulsifier E10 reagent Kit
•SOLiD™ EZ Bead™ Emulsifier E10 Accessories Kit
•SOLiD™ EZ Bead™ Enricher Buffer Kit
SOLiD™ EZ Bead™ E20 System Consumables, 4-pack (4453094)*
•SOLiD™ EZ Bead™ Emulsifier E20 reagent Kit
•SOLiD™ EZ Bead™ Emulsifier E20 Accessories Kit
•SOLiD™ EZ Bead™ Enricher Buffer Kit
SOLiD™ EZ Bead™ E80 System Consumables, 4-pack (4453095)*
•SOLiD™ EZ Bead™ Emulsifier E80 reagent Kit
•SOLiD™ EZ Bead™ Emulsifier E80 Accessories Kit
•SOLiD™ EZ Bead™ Enricher Buffer Kit
SOLiD™ EZ Bead™ Emulsifier Oil Pack Kit (4457185)SOLiD™ EZ Bead™ Emulsifier Oil Kit:
•SOLiD™ EZ Bead™ Emulsifier Oil Kit - Stabilizer 1
•SOLiD™ EZ Bead™ Emulsifier Oil Kit - Stabilizer 2
•SOLiD™ EZ Bead™ Emulsifier Oil Kit - Oil

SOLiD™ EZ Bead™ Emulsifier Oil Accessory Kit:

•SOLiD™ EZ Bead™ Emulsifier Oil Kit - Oil Mix Bottle
•SOLiD™ EZ Bead™ Emulsifier Oil Kit - Oil Funnel

 *The kit includes consumables for all three instruments of the SOLiD™ EZ Bead™ System:the SOLiD™ EZ Bead™ Emulsifier, SOLiD™ EZ Bead™ Amplifier and the SOLiD™ EZ Bead™ Enricher


[править] Before you begin

• Prepare an emulsion of the DNA libray and P1 Beads, either manually or automatically with the SOLiD™ EZ Bead™ Emulsifier
• Install the SOLiD™ EZ Bead™ Amplifier in a location to minimize environmental contamination to the sample.

[править] Prepare the SOLiD™ EZ Bead™ Amplifier Pouch

[править] Prepare the Oil Master Mix

  1. Unscrew and retain the cap with impeller of the SOLiD™ EZ Bead™ Emulsifier E-Large Jar.
  2. Tare the E-Large Jar on an analytical balance.
  3. Add each component to the E-Large Jar in the order below. Tare the E-Large Jar and contents before adding the next component:
    ComponentWeight (g)
    SOLiD™ EZ Bead™ Emulsifier Oil Kit - Stabilizer 1 11.2
    SOLiD™ EZ Bead™ Emulsifier Oil Kit - Stabilizer 2 2.7
    SOLiD™ EZ Bead™ Emulsifier Oil Kit - Oil 198.4

  4. Screw the cap with impeller on the E-Large Jar.
  5. Install the jar on the SOLiD™ EZ Bead™ Emulsifier:
    a. Turn the knob clockwise to open the compartment door.
    b. Pull the arm back at the base towards the opening of the compartment.
    c. Slide the jar until it stops against the two posts at the rear of the compartment.
    d. Allow the arm to rest against the jar (insert picture)
  6. Fit the impeller head into the locking slot. Slide the cuopler down until the cuopler clicks to a stop to ensure that the impeller head is firmly in place (insert picture)
  7. Start the run to mix the Oil Master Mix:
    a. Press the scale-selection button to select OIL on the instrument key panel
    b. Press the start button.

    The runtime is 5 minutes. after the run is complete, the SOLiD™ EZ Bead™ Emulsifier stops automatically, the right light on the panel is lit green, and the panel says, "Complete".

  8. Pour the prepared oil mix into provided SOLiD™ EZ Bead™ EmulsifierOil Kit-Oil Mix Bottle.
  9. Degas the Oil Master Mix for 30 minutes at 4°C.
  10. Store the Oil Master Mix at 4°C until use.

[править] Pre-rinse the SOLiD™ EZ Bead™ Amplifier Pouch

Pre-rinse the SOLiD™ EZ Bead™ Amplifier E-Small (for E10, E20) or E-Large (for E80) Pouch:

  1. Unscrew the cap from nozzle of the SOLiD™ EZ Bead™ Amplifier Pouch.
  2. Add an appropriate volume of the Oil Master Mix to the SOLiD™ EZ Bead™ Amplifier Pouch:
    ScalePouch to useVolume of Oil Master Mix to add(ml)
    E10, E20SOLiD™ EZ Bead™ Amplifier E-Small Pouch 10
    E80SOLiD™ EZ Bead™ Amplifier E-Large pouch 20

  3. Tightly screw the cap onto the nozzle of the SOLiD™ EZ Bead™ Amplifier Pouch to ensure that the pouch does not leak.
  4. Place the SOLiD™ EZ Bead™ Amplifier Pouch on the bench top. Massage the pouch for 2 to3 minutes to ensure that the Oil Master Mix coats the inner surface of the pouch.
  5. Over a waste container, unscrew the cap(s), then drain the Oil Master Mix into the container.
  6. Gently squeeze the SOLiD™ EZ Bead™ Amplifier Pouch to expel any remaining Oil Master Mix from the pouch.
  7. Drain the SOLiD™ EZ Bead™ Amplifier Pouch upside-down for at least 20 minutes on a layer of clean lint-free wipes. After drainig the pouch, the pouch is ready for use.

    NB! Do not re-use Oil Master Mix from pre-rinsing.

  8. Screw the cap(s) tightly on the nozzle(s) until SOLiD™ EZ Bead™ Amplifier Pouches are ready for use. Use the pre-rinsed SOLiD™ EZ Bead™ Amplifier Pouches the same day.

[править] Fill the pre-rinsed SOLiD™ EZ Bead™ Amplifier Pouch with emulsion

  1. Insert an empty, pre-rinsed SOLiD™ EZ Bead™ Amplifier Pouch into the slot(s) of the pouch filling station.
    ScalePouch to useSlot(s) to use
    E10, E20SOLiD™ EZ Bead™ Amplifier E-Small Pouch Middle slot
    E80SOLiD™ EZ Bead™ Amplifier E-Large pouch Outer slot

  2. Unscrew the cap(s) from the nozzle(s) of the pouch. TAke care not ton contaminate the cap(s).
  3. Fill the pouch with the emulsion:
    a. Insert the SOLiD™ EZ Bead™ Amplifier Funnel into a nozzle of the pouch.
    b. Unscrew the cap of the jar. Secure the impeller shaft with a finger against the cap, then tap excess emulsion from the impeller. Discard the cap.
    c. Hold the funnel firmly in place in the nozzle, then slowly pour all of the emulsion into the pouch. Let the emulsion drip from the jar for 30 seconds to recover as much of the emulsion as possible.
    d. Turn the crank of the pouch filling station counter-clockwise until most air is expelled from the pouch. Expelling ais from the pouch maintains the efficiency of the ePCR and ensures optimal filling of the pouch.
    e. Remove the funnel, screw the cap(s) onto the nozzle(s) until tight.
    f. Turn crank clockwise to release the pouch from the plates.

[править] Set up and run the SOLiD™ EZ Bead™ Amplifier

  1. Pull the lever all of the way until it stops to open the door to the SOLiD™ EZ Bead™ Amplifier.
  2. Remove the pouch frame from the SOLiD™ EZ Bead™ Amplifier, then place it on the bench top.
  3. Pull the foam away from the SOLiD™ EZ Bead™ Amplifier puoch frame to install the pouch nozzle(s) in the holder.
  4. Fit the bottom nozzle groove(s) of the pouch nozzle(s) into the slot(s) of the SOLiD™ EZ Bead™ Amplifier pouch holder. Install the single EZ Bead Amplifier E-Small Pouch into the middle slot of the Pouch Holder.
  5. Release the foam so that it returns to its original position in the frame.
  6. Place the pouch frame with installed pouch back on to the SOLiD™ EZ Bead™ Amplifier. Ensure that the pouch frame is properly seated on the Amplifier.
  7. Close the door to the SOLiD™ EZ Bead™ Amplifier and push the lever all of the way down until it stops to secure the door.
  8. Start the run:
    a. Open the SOLiD™ EZ Bead™ Amplifier software. Ensure that the control windows says "Ready"; and the green "POWER" on the front panel is lit. If the green "POWER" status is not lit, toggle the power switch to the on position on the back instrument panel, wait until the current run is complete.
    b. Click the scale-selection button to select the scale on the instrument key panel.
    c. Click the start-run button. The middle light in the control window is lit yellow, and the yellow "IN USE" on the front status panel is lit.

    The run time is ~2.5 hours (E10 scale) to ~3.5 hours (E20, E80 scales). After the run is complete, the SOLiD™ EZ Bead™ Amplifier stops automatically, and the control window of the software displays "Completed".

    NB! Do not open the door during the run. Opening the door compromises the sample because the sample is not be properly amplified. Stop the run, then prepare a new emulsion.
    The PCR plates are very hot.

  9. After the run is complete, remove the pouch from the SOLiD™ EZ Bead™ Amplifier and store the pouch at 4°C as soon as possible.

If the run is performed overnight or is left unattended on the SOLiD™ EZ Bead™ Amplifier after run completion, then the sample is stored automatically at 10°C. For best results, do not store the post-PCR sample on the Amplifier for >36 hours.

Stopping point
Store the post-PCR sample at 4°C or proceed to emulsion break and enrichment using the SOLiD™ EZ Bead™ Enricher.



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