genomic dna kit labchip gx/gxii user guide - bioneer gx... · genomic dna kit labchip gx/gxii user...

Caliper (a PerkinElmer company) of 21 Revised February 2013 68 Elm Street Hopkinton, MA 01748-1668 1-877-LABCHIP (522-2447)

Genomic DNA Kit

LabChip GX/GXII User Guide

Contents

SPECIFICATIONS ...................................................................................................................... 2

SAMPLE CONDITIONS .............................................................................................................. 2

REAGENT KIT AND CHIP.......................................................................................................... 3

SAFETY WARNINGS AND PRECAUTIONS .............................................................................. 4

PREPARATION PROCEDURES ................................................................................................ 5

ADDITIONAL ITEMS REQUIRED ........................................................................................ 5 PREPARING THE GEL-DYE SOLUTION ............................................................................. 5 PREPARING THE DNA SAMPLES, DNA LADDER AND BUFFER TUBE .................................. 6 PREPARING THE DNA CHIP ........................................................................................... 7 PREPARING THE LABCHIP GX INSTRUMENT .................................................................... 8 RUNNING THE GENOMIC DNA ASSAY ............................................................................. 8 STORING THE DNA CHIP ............................................................................................... 9

RESULTS ................................................................................................................................. 10

GENOMIC DNA SOFTWARE ANALYSIS .......................................................................... 10 GENOMIC DNA LADDER RESULT .................................................................................. 10 GENOMIC DNA RESULT .............................................................................................. 11

TROUBLESHOOTING .............................................................................................................. 12

LABCHIP KIT ESSENTIAL PRACTICES ................................................................................. 17

GENERAL ................................................................................................................... 17 REAGENTS ................................................................................................................. 17 CHIPS ........................................................................................................................ 18 SAMPLES .................................................................................................................... 19

CHIP WELL ASPIRATION USING A VACUUM ....................................................................... 20

CUSTOMER TECHNICAL SUPPORT ...................................................................................... 21

Genomic DNA LabChip Kit

LabChip® GX/GXII User Guide


Specifications1

Sizing Range 50 to 40,000+ bp

Sizing Accuracy ±20% Up to 10 kb, based on ladder

Sizing Precision 20% CV

Quantitation Range 2-50 ng/µL Sample diluted 10X with water

0.2-5 ng/µL Sample undiluted

Sensitivity 0.1 ng/µL Sample undiluted; S/N > 3; intact Human Control

gDNA

Carry-Over < 0.25%

Quantitation Accuracy ±30% Based on PicoGreen & plate reader quantitation of

Human Control gDNA

Quantitation Precision 20% CV Based on Human Control gDNA

Sample Volume Required 1 µL (diluted)

10 µL (undiluted) Requires 384-well plate

Samples per Chip Prep 48 or 24 Two workflows: one for ≤24 samples, one for ≤48

samples

Analysis Time 48 samples in 2.5 hrs Walk-away time

Samples per Chip Reagent Kit 480

Chip Reagent Kit Stability 3-9 months

Chip Lifetime 500 samples

Based on use with only one assay type for the lifetime of the chip; and under normal laboratory conditions and adherence to Caliper preparation protocols, sample guidelines and storage conditions. Individual results may vary.

1 Human Control gDNA from intestine was purchased from BioChain (Hayward, CA).

Sample Conditions

Additives

Caliper recommends that BSA and detergents exceeding 0.05 mg/mL and 0.01% v/v (respectively) in concentration not be used. Higher concentrations can result in chip failure. In addition, non-aqueous solvents are not compatible with DNA LabChip protocols.

Particulates All sample plates should be spun down prior to analysis. All buffers should be filtered

with a 0.22 µm cellulose acetate filter.

Salt Concentration Total salt concentration must not exceed 10 mM Tris. Higher salt concentrations and different ions may alter performance and reduce assay sensitivity.




Reagent Kit and Chip

Genomic DNA Reagent Kit Part Number CLS760685 Kit contains enough reagents for 20 Small-batch or 10 Large-batch chip preparations. Up to 24 samples can be tested with a Small-batch chip preparation. Up to 48 samples can be tested with a Large-batch chip preparation.

Reagents Label Quantity

DNA Dye Concentrate Blue 1 vial, 0.09 mL

Chip Storage Buffer White 9 vials, 1.8 mL each

Genomic DNA Gel Matrix Red 5 vials, 1.1 mL each

10X Genomic DNA Ladder Yellow 1 vial, 0.26 mL

Genomic DNA Marker Green 1 vial, 1.5 mL

Consumable Items Supplier Number Quantity

Spin Filters Costar, Cat. # 8161 10

Ladder Tubes Genemate, Cat. # C-3258-1 20, 0.2 mL

Detection Window Cleaning Cloth VWR, Cat. # 21912-046 1

Swab ITW Texwipe®, Cat. # TX758B 3

Buffer Tubes E&K Scientific, Cat. # 697075-NC 20, 0.75 mL

HT DNA Extended Range Chip Part Number 760517

Item Quantity

DNA Chip 1




Safety Warnings and Precautions ! WARNING ! For Research Use Only. Not recommended or intended for diagnosis of disease in humans or animals. Do not use internally or externally in humans or animals. CAUTION We recommend that this product and components be handled only by those who have been trained in laboratory techniques and that it is used in accordance with the principles of good laboratory practice. As all chemicals should be considered as potentially hazardous, it is advisable when handling chemical reagents to wear suitable protective clothing, such as laboratory overalls, safety glasses, and gloves. Care should be taken to avoid contact with skin or eyes. In case of contact with skin or eyes, wash immediately with water. ! WARNING ! Dye Concentrate contains DMSO. S24/25: Avoid contact with skin and eyes.

Storage: When not in use, store chips and reagents refrigerated at 4 °C. Do not leave chips and reagents unrefrigerated overnight.




Preparation Procedures

Additional Items Required

• MilliQ water: Molecular biology grade or better, 0.22-micron filtered

• 70%-isopropanol solution in DI water

• Bio-Rad Hard-Shell® 384-well Skirted PCR Plates, catalog # HSP-38XX (recommended)

• Bio-Rad Hard-Shell® 96-well Skirted PCR Plates, catalog # HSP-96XX (recommended)

Preparing the Gel-Dye Solution

1. Gently vortex the thawed DNA Dye Concentrate (blue cap ) for 10 seconds before use.

2. Transfer 13.75 µµµµL DNA Dye Concentrate (blue cap ) to 1 vial of Genomic DNA Gel Matrix (red cap ).

3. Vortex the solution until it is well mixed and then spin down the mixture for a few seconds.

4. Transfer the mixture into two spin filters (approximately 550 µL each).

5. Centrifuge at 9200 rcf for 7.5 minutes at room temperature. Store in the dark at 4 °C. Use within 3

weeks.

Note: The Dye Concentrate contains DMSO and must be thawed completely before use.

One vial of Genomic DNA Gel Matrix (red cap ) is good for 4 Small -batch or 2

Large-batch chip preparations. Up to 24 samples can be tested with a Small-batch chip preparation. Up to 48 samples can be tested with a Large-batch chip preparation.

Note: Allow the chip and reagents to equilibrate to room temperature for about 20 minutes before use.




Preparing the DNA Samples, DNA Ladder and Buffer Tube

1. In the provided 0.2 mL Ladder Tube, add 12 µµµµL of Genomic DNA Ladder (yellow cap ) to 108 µµµµL

molecular biology-grade water. Mix thoroughly by pipetting the solution up and down several times. (Note:

Do not vortex ladder solution. Vortexing may degrade large DNA ladder fragments.)

2. Insert the Ladder Tube into the ladder slot on the LabChip GX instrument.

3. Prepare samples in a 384-well plate by adding 2 µµµµL of sample to 18 µµµµL of molecular biology-grade water.

(Note: If sample volume is limited, 1 µµµµL of sample can be added to 9 µµµµL of molecular biology-grade water.

However, due to evaporation, samples can only be tested in Small-batch runs, ≤ 24 samples, and using a

384-well sample plate. Additionally, samples at concentrations from 0.2 to 5 ng/µL should be run undiluted.

The reported LabChip GX concentration must then be divided by 10 to account for the omission of the 10X

dilution.)

4. Add 750 µL molecular biology-grade water to the 0.75 mL Buffer Tube provided with the reagent kit.

5. Insert the Buffer Tube into the buffer slot on the LabChip GX instrument.

Buffer Tube

Ladder Tube

384-well Sample Plate

96-well Sample Plate (requires 2+18 µL workflow and low sip)

Maximum number of samples per run = 48

Sample Preparation

Recommended: 2 µL sample + 18 µL water

or

Limited Sample: 1 µL sample + 9 µL water Not for Large-batch runs (>24 samples)

Samples at concentrations from 0.2 to 5 ng/µL should be run undiluted; reported LabChip GX

concentration must then be divided by 10.

750 µL molecular biology-grade water

1X Ladder =

12 µL DNA Ladder + 108 µL water Note: Do not vortex ladder solution. Vortexing may degrade large DNA ladder fragments.

LabChip GX

Figure 1. Sample Workflow




Preparing the DNA Chip

1. Allow the chip to come to room temperature.

2. Use a pipette tip attached to a vacuum line to thoroughly aspirate

all fluid from the chip wells (see Figure 2). For more details on

how to setup a vacuum line see page 20.

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

and 10) twice with molecular biology grade water. Do not allow

active wells to remain dry.

4. If any water spilled onto on the top and bottom chip surfaces

during rinsing, aspirate using vacuum line. DO NOT run the tip

over the central region of the detection window.

5. Using a Reverse Pipetting Technique, add Gel-Dye to chip well 3,

7, 8 and 10 as shown in Figure 3. For Small-batch add 50 µL per

well. For Large-batch add 75 µL in wells 3, 7 and 8 and 120 µL

in well 10.

6. Add Genomic DNA Marker (green cap ) to chip well 4 as shown

in Figure 3. For Small-batch add 60 µL. For Large-batch add

120 µL. (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.)

7. Ensure chip well 1 (or waste well) is empty before placing the chip

on the LabChip GX.

Figure 2. Using a vacuum to aspirate the chip wells is more effective than using a pipette.

Figure 3. Chip Preparation




Preparing the LabChip GX Instrument

1. Place the sample plate, Ladder Tube and

Buffer Tube onto the plate holder of the

LabChip GX.

2. Remove the chip from the chip storage

container and inspect the chip window.

Clean BOTH sides of the chip window with

the supplied cleanroom cloth dampened with

a 70%-isopropanol solution in DI water.

3. Eject the chip cartridge by pressing the CHIP

button on the instrument front panel.

4. Release the cartridge latch, insert the chip into

the LabChip GX instrument, refasten the latch and push the cartridge into the instrument.

5. The Select Assay dialog box will pop up. Select the HT gDNA assay. (Note: The chip may be run with

multiple assays, but only one assay type should be run on the chip.)

6. Press the EJECT button on the instrument front panel to retract the sample plate and send the Buffer

Tube to the sipper.

Running the Genomic DNA Assay

1. Start the LabChip GX software.

2. On the main screen, click on the Run button in the upper left corner of the LabChip GX software.

3. The Start Run dialog box will pop up with tabs listed as Output, Run and Advanced.

4. In the Run Tab, select assay type HT gDNA and the appropriate Operator Name, Plate Name, well

pattern and barcode option.

5. In the Output Tab select the destination of the file, the filename convention and any additional data to

export automatically.

6. In the Advanced Tab, select the number of times each well is sampled, the inclusion of any sample names

and any expected peaks.

7. Click Start to begin the run.

Ladder Tube

Buffer Tube

Sample Well A1

Note: If performing multiple runs in a day, the chip should be washed in between chip preparations, using the instrument and Chip Storage buffer as described in the “Storing the DNA Chip” section.

Be sure to periodically clean the O-rings on the top plate of the chip interface on the LabChip GX. With the provided lint free swab dampened with DI water, clean the O-rings using a circular motion. Allow the O-rings to dry before inserting a chip.

Figure 4. LabChip GX Plate Tray




Storing the DNA Chip After use, the chip must be cleaned and stored in the chip container. The procedure below can be conducted the

following day when running overnight.

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

2. Each active well (1, 3, 4, 7, 8, and 10) should be rinsed and aspirated twice, using molecular biology grade

water.

3. Add 100 µL of DNA Chip Storage Buffer (white cap ) to the active wells.

4. Place the chip back on the LabChip GX. Ensure a Buffer Tube with 750 µL sample buffer or molecular

biology-grade water is in the buffer slot and click the Wash button.

5. Remove the chip from the instrument and place in storage container.

6. Add an additional 50 µL of Chip Storage Buffer to wells 1 and 4.

7. Cover the wells with Parafilm® to prevent buffer evaporation and store at 4 °C. Storage of a chip with dry

wells may cause it to become clogged.

Chip Cartridge Cleaning

1. Daily

A) Inspect the inside of the chip cartridge and O-rings for debris.

B) With the provided lint free swab dampened with DI water, clean the O-rings using a circular motion.

If the O-rings stick to the chip or a pressure leak is detected, perform the more extensive monthly

cleaning procedure.

2. Monthly

A) To reduce pressure leaks at the chip interface, clean the O-rings frequently. Remove the O-rings

from the top plate of the chip interface on the LabChip GX instrument. Soak O-rings in DI water for

a few minutes. Clean the O-ring faces by rubbing between two gloved fingers.

B) To reduce the occurrence of current leaks, clean the chip interface frequently. Clean the top plate of

the chip interface using the provided lint free swab dampened with DI water.

C) Allow the O-rings and chip interface to air dry. Reinsert the O-rings into the chip cartridge.




Results

Genomic DNA Software Analysis Data are analyzed by aligning the sample data and normalizing the sample area, using the Lower Markers in the

samples and in the two ladders that bracket every 12 samples. The size of a sample is determined by comparing the

migration times of peaks within the sample to those of the fragments of known size in the bracketing ladders. The

concentration of a sample is calculated using a Total Smear, starting at 0.18 kb and extending to 300 kb (see Figure

5). These smear limits can be adjusted by the user. A calibration curve generated using the known size and

concentrations of ladder peaks is applied to the normalized area of this Total Smear, to determine the concentration

of the sample. This Total Smear Concentration is reported in the Well Table entry of each sample.

The Genomic DNA assay also reports a Genomic DNA Quality Score (GQS) in the Well Table entry of each sample.

This score represents the degree of degradation of a sample, with 5 corresponding to intact gDNA and 0

corresponding to highly degraded gDNA, and is calculated from the size distribution of a sample.

Figure 5. Genomic DNA Total Smear (in orange).

Genomic DNA Ladder Result

The electropherogram of a typical Genomic DNA ladder is shown in

Figure 6. Following the Lower Marker are ladder fragments of 100, 300, 500, 700, 1100, 1900, 2900, 4900, 7000,

10000 and 40000 bp.




Figure 6. Genomic DNA Ladder

Genomic DNA Result

An electropherogram and gel view of intact and degraded genomic DNA from BioChain is shown in Figure 7.

Genomic DNA was degraded by incubating with Fragmentase® for 10 or 20 minutes. Genomic DNA treated for 10

minutes was partially degraded (GQS = 2.5), while genomic DNA treated for 20 minutes was highly degraded (GQS =

0).

Figure 7. Intact and degraded genomic DNA from BioChain.

Lower Marker




Troubleshooting

1. Symptom: No ladder or sample peaks but Lower Marker peak detected. The Lower Marker peak height will most likely be greater than normal height.

Figure 8. Electropherogram with no ladder or sample peaks, but lower marker peak is present

A) During chip priming, an air bubble in the sipper may have been introduced. To remove the air bubble,

reprime the chip. See the section entitled “LabChip Kit Essential Practices – Chips” for instructions on

how to reprime the chip.

B) If the air bubble is not dislodged after a reprime, then apply a vacuum to the sipper. Perform this by

filling all active wells with 100 µL of Chip Storage Buffer. Then suction the sipper with a vacuum line, as

shown in Figure 9, until droplets of fluid flow out from the sipper. When suctioning the sipper be careful

not to bend or break the sipper. To facilitate this, cut the tip of the vacuum line to widen the mouth.

Lower Marker




Figure 9. Removing an air bubble or clog by suctioning the sipper with a vacuum line

2. Symptom: Ladder detected but no sample peaks

A) The sipper may not be reaching the sample due to low sample volume in the well plate. Add more

sample to the well or dilute sample with 1X TE buffer.

B) If the missing sample peaks occurred only in a few wells of the plate, check those wells for air bubbles.

If present, remove air bubbles by manually inserting a larger volume pipette tip (~100 µL) into the

sample well and dislodging the bubble. Then retest these sample wells.

C) The sipper may not be reaching the sample due to an incorrect capillary height setting or misalignment

between the sipper and the wells of the sample plate. To correct the former, try the low capillary height

setting in the Run menu. To correct the latter, re-teach the robot positioning as described in the LabChip

GX User’s Manual.

D) If the plate has been uncovered for some time, sample may have evaporated. Check the sample wells,

especially near the edge of the plate where evaporation is fastest, and replenish wells with insufficient

volume by adding or diluting samples.

E) Debris from the sample or sample preparation is clogging the sipper. If you suspect there may be

debris, spin the sample plate down in a centrifuge. Unclog the sipper by repriming the chip or

vacuuming the sipper as described in Troubleshooting Item #1.




3. Symptom: No ladder peaks but sample peaks and marker peak are present.

A) There may be low or no ladder volume in Ladder Tube. Add more ladder (diluted as described in the Preparing the DNA Samples, DNA Ladder and Buffer Tube section) to the Ladder Tube and restart the run. The recommended minimum ladder volume is 100 µµµµL.

4. Symptom: No Lower Marker peak but sample peaks are present.

A) Insufficient or no marker added to chip well 4. The marker volume may also become too low if the chip

remains idle on instrument for extended period of time due to gravity-driven flow. Add or replenish the

marker solution in the chip.

B) If there is marker solution in well 4, the problem may be due to a marker channel clog. Unclog the

marker channel by repriming the chip or vacuuming the sipper as described in Troubleshooting Item

#1.

5. Symptom: Peaks migrating much faster than expected.

A) Incorrect Gel-Dye concentration. Migration time is sensitive to dye concentration and peaks will

migrate too fast or too slow if the dye concentration in the gel is too low or too high, respectively.

Prepare a fresh Gel-Dye solution. Wash and reprime the chip with the new Gel-Dye mixture.

B) If fast migration is observed repeatedly on a new chip then contact technical support.

NOTE: Some migration time variance between chips or within a plate is considered normal chip performance. All chips are QC tested at Caliper Life Sciences prior to shipment.




6. Symptom: Peaks migrating much slower than expected.

An electropherogram of a genomic DNA sample is shown in

Figure 10 where the migration time is much longer than expected.

Figure 10. Electropherogram of genomic DNA migration that is too slow.

A) Particulates from the samples may be clogging the separation channel. Minimize the loading of

particulates in the sample by spinning down the plate (e.g. 3000 rcf for 5 mins) before testing and/or

selecting a plate type with a higher sip height in the Start Run dialog box before starting a new run.

The debris maybe flushed out of the chip by washing and repriming the chip.

B) Excess dye within the separation channel. Prepare a fresh Gel-Dye solution. Wash the chip and then

reprime with the new Gel-Dye mixture.

C) Gel-Dye was not primed properly into the chip. Check the O-rings on the top surface of the chip

interface and clean if necessary. Then reprime the chip.

7. Symptom: Unexpected sharp peaks.

Electropherograms of genomic DNA are shown in Figure 11 and Figure 12 with unexpected sharp peaks

caused by particulates (like dust) or aggregates of large DNA that can form during sample loading and/or

migration in the chip.

A) Dust or other particulates introduced through sample or reagents. Replace molecular biology-grade

water used for chip preparation. Replace buffer used for sample and reagent preparation. Use a

0.22-micron filter for all water and buffers used for chip, sample, and reagent preparation.

Lower Marker

genomic DNA




B) Aggregates of large genomic DNA. Retest sample. Dilute sample further, if possible, to reduce

aggregation.

Figure 11. Electropherogram with an unexpected peak migrating after a genomic DNA sample

Figure 12. Electropherogram with an unexpected peak that overlaps with the migration of a genomic DNA sample

Lower Marker

Lower Marker

Peak caused by particulates or DNA aggregates.

Peak caused by particulates or

DNA aggregates.




LabChip Kit Essential Practices To ensure proper assay performance please follow the important handling practices described below. Failure to observe these guidelines may void the LabChip Kit product warranty.

1

NOTE: It is important to keep particulates out of the chip wells, channels and capillary. Many of the following guidelines are designed to keep the chips particulate-free. For assay and instrument troubleshooting, refer to the LabChip GX Software Help file or call Caliper Technical Support at 1-877-LABCHIP.

General

• Allow the chip, sample plate and all reagents to equilibrate to room temperature before use (approximately

20 to 30 minutes).

• Clean the O-rings in the chip interface weekly and the electrodes daily. Refer to the Instrument User

Guide Maintenance and Service section for procedures.

• Avoid use of powdered gloves. Use only non-powdered gloves when handling chips, reagents, sample

plates, and when cleaning the instrument electrodes and electrode block.

• Calibrate laboratory pipettes regularly to ensure proper reagent dispensing.

• Only the Caliper-supplied clean room cloth can be used on the chip to clean the detection window. Use of

other, non-approved wipes may leave fluorescent debris, which can cause erratic focusing.

• Water used for chip preparation procedures must be 0.22-micron filtered, deionized, molecular biology-

grade.

• Use of the “Reverse Pipetting Technique” (described below) will help avoid introducing bubbles into the

chip when pipetting gel or other viscous solutions.

Reverse Pipetting Technique Step 1. Depress the pipette plunger to the second stop. Step 2. Aspirate the selected volume plus an excess amount from

the tube. Step 3. Dispense the selected volume into the corner of the well by

depressing plunger to the first stop. Step 4. Withdraw the pipette from the well.

Reagents

• Store all reagents at 4 °C when not in use.

• The LabChip dye contains DMSO and should be thawed completely before use. It is recommended that

you prepare aliquots to reduce the time required for thawing.

• Gently vortex all kit reagents.

• Dispense reagents into chip wells slowly, without introducing air bubbles. Insert the pipette tip vertically

and to the bottom of the chip well.

• Protect the dye and Gel-Dye mixture from light. Store in the dark at 4 °C when not in use.

1 Caliper (a PerkinElmer company) warrants that the LabChip Kit meets specification at the time of shipment, and is free from defects in material and

workmanship. LabChip® Kits are warranted for 90 days from the date of shipment. All claims under this warranty must be made within thirty days of the discovery of the defect.




Chips

• Repriming Chips:

� Press the CHIP button on the front instrument panel to eject the chip cartridge.

� Reinsert the cartridge by pushing the cartridge back into the instrument.

� Click the Run button on the main screen of the LabChip GX software.

• Washing and Repriming Chips:

� Press the CHIP button on the front instrument panel to eject the chip cartridge.

• Open the chip cartridge and return the chip to the chip container, ensuring the sipper is

immersed in fluid.

• Thoroughly aspirate all fluid from the chip wells using a vacuum line.

• Ensure that each active well (1, 3, 4, 7, 8 and 10) is rinsed and completely aspirated twice

with molecular biology-grade water. Do not allow active wells to remain dry.

• Add 100 µL of Chip Storage Buffer to each active well (1, 3, 4, 7, 8 and 10).

• Place the chip in the LabChip GX instrument.

• Reinsert the cartridge by engaging the latch and pushing the cartridge back into the

instrument.

• Click the Wash button on the main screen of the LabChip GX software.

� After completion of the wash cycle, Press the CHIP button on the front instrument panel

to eject the chip cartridge.

• Open the chip cartridge and return the chip to the chip container, ensuring the sipper is

immersed in fluid.

• Thoroughly aspirate all fluid from the chip wells using a vacuum line.

• Ensure that each active well (1, 3, 4, 7, 8 and 10) is rinsed and completely aspirated twice

with molecular biology-grade water. Do not allow active wells to remain dry.

• Prepare chip as described in the “Preparing the DNA Chip” section.

• Place the chip in the LabChip GX instrument.

• Reinsert the cartridge by engaging the latch and pushing the cartridge back into the

instrument.

• Click the Run button on the main screen of the LabChip GX software.

• Other Considerations:

• When not in use the active wells should be covered with Parafilm® and the chip should

be stored at 2-8 °C.

• If using the chip again within 24 hours, it may be left at room temperature.

• Do not allow the liquid in the chip container to freeze, as this may lead to poor chip

performance. Do not submerge the chip in any solution.

• The entire chip surface must be thoroughly dry before use.




• The sipper must be kept immersed in fluid at all times and should not be exposed to air

for long periods of time.

• Use care in chip handling to prevent sipper damage. Damage to the sipper can result in

inconsistent sampling.

• Avoid exposing the chips to dust by keeping them in a closed environment, such as in

the chip container or in the instrument, before and after chip preparation.

• Chips can be prepared and left idle on the instrument for up to 8 hours. This workflow

allows analysis of samples as needed throughout the day, without having to re-prep the

chip, as long as the maximum number of samples per chip prep is not exceeded.

Samples

• Prepared sample plates should be free of gas bubbles and particulate debris, both of which may inhibit

sipper flow.

• Sample plates containing gas bubbles and/or particulate debris should be spun down (for example, 3000

rcf for 5 mins at RT) prior to analysis.

‘




Chip Well Aspiration Using a Vacuum

Aspirating with a pipette can leave used reagents in the chip wells. For this reason, Caliper recommends vacuuming the wells instead. This can be accomplished by attaching a permanent pipette tip to a house vacuum line with trap (Figures 1a and 1b). To avoid contamination, use a new pipette tip over the permanent tip for each chip aspirated (Figure 2).

Figure 1a Figure 1b

Figure 2




Customer Technical Support

Caliper (a PerkinElmer company) 68 Elm Street Hopkinton, MA 01748-1668 Phone: 1-877-LABCHIP (522-2447) Fax: 1-508-435-3439 For additional assay and instrument troubleshooting, refer to the LabChip GX Software Help file or call Caliper Technical Support at 1-877-LABCHIP.

The chip and reagents supplied with this kit are sold with limited rights of use. The chip may only be used with the specific quantity of reagents supplied with this kit. The purchaser has no right or license to refurbish, reuse, remanufacture, or otherwise use the chip with any other reagents than those specifically supplied in this kit. For more information on the terms and conditions of use of these chips and reagents, please read your LabChip® GX User Guide. Caliper, the Caliper logo, LabChip, and the LabChip logo are registered trademarks of Caliper (a PerkinElmer company). The reagent kits contain materials manufactured for Caliper by Molecular Probes, Inc., and are provided under a license from Molecular Probes, Inc., for only use in Research, Human Diagnostics, Biohazard Detection, Environmental Testing, Food Testing, Quality Control, and Pathogen Testing. © Copyright Caliper (a PerkinElmer Company) 2013 http://www.caliperLS.com Doc CLS136856

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