RNA Assay Quick Guide LabChip GX Touch GXII Touch说

2024-07-28 作者:

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Allow the chip and refrigerated reagents to equilibrate to room temperature for at least 30 minutes

before use. Thaw the RNA Ladder on ice.

RNA Dye contains DMSO and must be thawed completely before use.

The dye is light sensitive. Do not expose the Dye or Gel-Dye solution to light for any length of time.

Keep the prepared Gel-Dye solution in the dark. Preparation of Gel-Dye Solution

1. Vortex the thawed RNA Dye Concentrate for 10 to 15 seconds before use.

2. Transfer 75 µL of RNA Dye Concentrate to a 2.0 mL centrifuge tube provided with the reagent kit.

3. Add 425 µL of RNA Gel Matrix using a Reverse Pipetting Technique.

4. Vortex the Gel-Dye solution until it is well mixed and spin down for a few seconds.

5. Transfer the Gel-Dye solution to a spin filter. Use a centrifuge tube filled with 500 µL of water

(Milli-Q® or equivalent) to balance the centrifuge.

6. Centrifuge at 9300 rcf for 10 minutes at room temperature.

7. Discard the filter.

8. Label and date the tube.

9. Store in the dark at 2-8°C. Use within 5 days.

Low-Throughput (LT) Chip Preparation, up to 48 samples

High Throughput (HT) Chip Preparation, up to 192 samples

1. Rinse and completely aspirate each active well (1, 3, 4, 7, 8, and 10)

twice with water (Milli-Q® or equivalent). Do not allow the active wells to

remain dry.

2. If any water spills onto the top or bottom chip surfaces during rinsing,

aspirate using the vacuum line. DO NOT run the tip over the detection window. Use the provided Detection Window Cleaning Cloth

dampened in water (Milli-Q® or equivalent) or 70% isopropanol to

clean the detection window as needed.

3. Using a Reverse Pipetting Technique, add Gel-Dye solution to chip wells

3, 7, 8, and 10 as shown in Figure 1 (LT)

or

Figure 2 (HT).

4.

Add

50

μL (LT) or

100 μl (HT) RNA Marker to chip well 4 as shown in

Figure 1 (LT) or Figure 2 (HT).

Note: The marker well may need to be replenished if the chip is in idle mode

on the instrument for an extended period of time.

5. Make sure the rims of the chip wells are clean and dry.

6. IMPORTANT: Ensure chip well 1 (waste well) is empty before placing

the chip into the LabChip GX Touch/GXII Touch.

Figure 2. High-throughput (HT)

chip preparation

Figure 1. Low-throughput (LT)

chip preparation

RNA Sample, Ladder, and Buffer Preparation

1. Prepare 1X Sample Buffer by adding 620 μL RNA Sample Buffer Concentrate to 5580 μL

DEPC treated or nuclease-free water.

Note: The RNA Sample Buffer Concentrate is a 10X solution. Sample Buffer is stable after dilution, but to avoid RNase contamination, sample buffer should be prepared fresh.

2. Allow the RNA ladder to thaw on ice. (Avoid multiple freeze/thaws. It is recommended to aliquot the

RNA ladder into five 4 µL lots for individual use after thawing the vial for the first time.)

3. Transfer 4 μL RNA Ladder

into the provided 0.2 mL Ladder Tube and cover.

4. For each sample to be analyzed, pipette 2 μL (RNA Std Sens) or 6 μL (RNA High Sens) sample

into individual microtiter plate wells. Cover with PCR cap strips.

5. Heat the ladder and samples at 70°C for 2 minutes.

6. Snap cool the samples and ladder by immediately placing the tubes and/or

microtiter plate on ice for 5 minutes.

7. Add 46 μL (RNA Std Sens) or 19 μL (RNA High Sens) prepared 1X Sample

Buffer to each sample. Mix by pipetting up and down a few times. Avoid

creating air bubbles. Cover the samples with PCR strip caps and spin down

the plate at 3000 rpm for 5 minutes.

8. Add 96 μL prepared 1X Sample Buffer to the Ladder Tube.

9. Add 750 μL prepared 1X Sample Buffer to the provided Buffer Tube.

Chip Cleaning and Storage

After use, the chip must be cleaned and stored in the chip container.

1. Place the chip into the chip storage container. Verify the sipper is submerged in the fluid reservoir.

2. Remove the reagents from each well of the chip using a vacuum.

3. Rinse and completely aspirate each active well (1, 3, 4, 7, 8, and 10) twice with water (Milli-Q® or

equivalent).

4. Add 120 μL of RNA Chip Storage Buffer (white cap

6. Touch the Wash button on the Home screen.

7. Touch the Wash button on the Wash screen.

8. When the chip wash is complete, remove the chip from the instrument and place the chip into the chip

storage container.

9. Add an additional 50 μL RNA Chip Storage Buffer to well 1.

10. Cover the wells with Parafilm® to prevent evaporation and store at 2-8°C. Storing a chip with dry wells

may clog the chip. If using the chip again within 24 hours, the chip can be stored at room

temperature.

) to the active wells.

5. Place the chip back into the LabChip GX Touch/GXII Touch.

Assay Specifications

The RNA Assay is for use with LabChip GX Touch/GXII Touch instruments. LabChip GX Touch/GXII

Touch instruments are for research use only and not for use in diagnostic procedures. Linear Range

Quantitation Reproducibility Size Range

RNA Sample Volume

Run Time Setup Time

Samples per Chip Prep

Chip Preps per

Reagent Kit Chip Lifetime

25 ng/ μL – 250 ng/ μL (Std Sens)

5 ng/ μL – 50 ng/ μL (High Sens) 20% CV

100 – 6000 nucleotides (suitable for total RNA)

2 μL of user sample (Std Sens)

6 μL of user sample (High Sens) 80 seconds per sample

(about 2.5 hours for 96 samples)

Approximately 30 minutes to prepare chip and samples

Up to 192 samples per HT chip prep

Up to 48 samples per LT chip prep

5 HT chip preps or

10 LT chip preps

HT: 2000 samples 24: 750 samples

Contact PerkinElmer

PerkinElmer, Inc. 68 Elm Street, Hopkinton, MA 01748-1668 USA

PerkinElmer Technical Support

Phone: (USA Toll Free) 800-762-4000; (Worldwide) +1 203-925-4602

Email: *********************************

For the complete RNA Assay User Guide, go to: /

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