DxDirect Gene Expression User Manual ver 1

DxDirect Gene Expression User Manual ver 1
DxDirectTM: Multiplexed Gene Expression
Analysis Direct from DxCollect RNA Stabilized
Blood
Compatible with DxTerity DxCollectTM Blood RNA Collection Tubes
User Manual
1
For Research Use Only. Not for Use in Diagnostic Procedures
For Research Use Only. Not for Use in Diagnostic Procedures.
Information in this document is subject to change without notice.
Limited License
Subject to the DxTerity terms and conditions that govern your use of DxTerity products, DxTerity grants you a nonexclusive,
nontransferable, nonsublicensable license to use this DxTerity product only in accordance with the manual and written
instructions provided by DxTerity with no rights for resale, commercial use, or reformulation. You understand and agree that,
except as expressly set forth in the DxTerity terms and conditions; no right or license to any patent or other intellectual
property owned or licensable by DxTerity is conveyed or implied by this DxTerity product. In particular, no right or license is
conveyed or implied to use this DxTerity product in combination with a product not provided, licensed, or specifically
recommended by DxTerity for such use. The license provided specifically excludes use for diagnostic applications.
Trademarks
DxTerity, DxDirect, and DxCollect are registered trademarks of DxTerity Diagnostics, Inc.
Applied Biosystems, 3500, 3500XL, 3730, and 3730XL are registered trademarks of Thermo Fisher, Inc.
All other trademarks are the property of their respective owners.
Citing DxDirect and DxCollect in Publications
When describing a procedure for publication using this product, please refer to it as the DxDirect Gene Expression Assay from
DxTerity.
Copyright
2014 DxTerity Diagnostics, Inc. All rights reserved
2
For Research Use Only. Not for Use in Diagnostic Procedures
Table of Contents
Use Limitations ..................................................................................................................................4
DxDirect Reagent System Contents and Storage Conditions ................................................................4
Equipment and Reagents Required but not Supplied...........................................................................5
Recommended Capillary Electrophoresis Platforms ............................................................................5
Technical Support ..............................................................................................................................5
Introduction ......................................................................................................................................6
How DxDirect Works ..........................................................................................................................6
Detailed Procedure ............................................................................................................................9
1. Sample Preparation............................................................................................................................... 9
2. Hybridization and Ligation .................................................................................................................... 9
3. Target Capture .................................................................................................................................... 10
4. Amplification ....................................................................................................................................... 10
5. Detection............................................................................................................................................. 11
DxDirect Assay Worksheet: Ligation and Target Capture ................................................................... 13
DxDirect Assay Worksheet: Amplification Setup ............................................................................... 14
Appendix A ...................................................................................................................................... 15
Contact Information......................................................................................................................... 17
3
For Research Use Only. Not for Use in Diagnostic Procedures
Use Limitations
The DxDirect reagents are intended for Research Use Only.
DxDirect Reagent System Contents and Storage Conditions
The components of the DxDirect Reagent System and their recommended storage conditions are listed
below. The DxDirect Reagent System is designed for use with magnetic plate washers and is available in
2 sizes—200 reactions and 1,000 reactions. Kit components have a shelf life of 6 months from date of
receipt.
Store reagents according to labels on boxes.
Do not pool reagents or mix and match kit components from different lots.
Reagent Box 1: Store between -30C and -15C
Description
Size
Storage
DirectPrime—Universal amplification primers for up to 40 targets
multiplexed assays
1 mL
-30C to -15C
DirectTaq—PCR Reaction Mix containing enzyme for 200 reactions
1 mL
-30C to -15C
Description
Size
Storage
DirectMix C—Proteinase K for 200 reactions
1 mL
2C to 8C
DirectBeads—Magnetic capture beads for 200 reactions
2 mL
2C to 8C
DirectReact—Reaction buffer for 200 reactions
2 mL
2C to 8C
Size
Storage
2 x 125 mL
2C to 25C
Reagent Box 2: Store between 2C and 8C
DirectWash: Store between 2 to 25C
Description
DirectWash—Wash buffer for 200 reactions
4
For Research Use Only. Not for Use in Diagnostic Procedures
Equipment and Reagents Required
Required Material and Equipment
Recommended
DirectMix A
User supplied
DirectMix B
User supplied
PCR Thermal Cyclers with Hot Lids
PCR Thermal Cycler with a temperature range from 4.0°C to
99.9°C; maximum block ramp rate of 3.9°C⁄sec
Vortex Mixer
Any model
Centrifuge for 96-well Plates
Any model
Adjustable Volume Pipettes (single
and 8-channel)
Any model
Handheld Magnetic Capture Plate
Life Technologies DynaMag™—96-well Side-Skirted Magnetic
Plate (CAT # 120-27)
Lab Consumables
96-well 0.2 mL PCR plates; 8-well strip covers or plate tape
seals; and reagent reservoirs
Recommended Capillary Electrophoresis Platforms
Polymer
Formulation
Model
Array
Applied Biosystems
(ABI) 3130
ABI 3500 and
3500XL
8- & 16-capillary
array
8- & 24-capillary
array
ABI 3730
48-capillary array
POP-6 & POP-7
ABI 3730XL
96-capillary array
POP-6 & POP-7
POP-6 & POP-7
POP-6 & POP-7
Technical Support
For questions regarding the DxDirect Assay, contact:
DxTerity Technical Support
19500 S. Rancho Way, Suite 116
Rancho Dominguez, CA 90220
Telephone: +1 310.537.7857
e-mail: [email protected]
5
For Research Use Only. Not for Use in Diagnostic Procedures
Introduction
The DxDirect assay technology combines Chemical Ligation Probe Amplification (CLPA) technology with
a proprietary blood stabilization buffer, DxCollect, to enable direct from sample analysis of genomic
signatures. CLPA uses a robust, non-enzymatic chemical reagent system to enable the direct
quantitative analysis of RNA and DNA in denaturing chemical environments like those experienced in
stabilized, unpurified blood. Sample processing and multiplexed, end-point PCR is performed in a single
reaction tube, and the final assay amplicon detection is accomplished using capillary electrophoresis
instruments.
How DxDirect Works
DxDirect must be used in conjunction with probes designed by the user according to the DxDirect
Probe Design User Manual and probe mixtures optimized according to the Assay Optimization
Application Note
For every gene in the assay, a unique probe set is designed
consisting of two DNA oligonucleotides, known as the Thiol Group
Probe (S-Probe) and the Leaving Group Probe (L-Probe), that bind
adjacently to one another and capture probes upstream or
downstream from the target site (refer to Figure 1). Each S- and LProbe pair is designed to contain a target-specific hybridization
sequence and a unique identification sequence that enables size
separation by CE. All S- and L-Probes contain upstream and
downstream universal primer sequences to allow for multiplex
PCR amplification of all ligation products.
In the DxDirect method (refer to Figure 2), S- and L-Probes
hybridize adjacently to one another leading to a chemical
LIGATION reaction triggered by proximity of the Leaving (L) group
to the Thiol (S) group. The ligation products are generated directly
in the denatured sample; CAPTURED on magnetic beads to
remove un-reacted probes; and then AMPLIFIED by PCR directly
from the beads. The final amplified product is DETECTED by CE,
based on final amplicon size. The assay is quantitative because the
amount of each ligation product made during the DxDirect
reaction is proportional to the concentration of its RNA target
sequence, and the relative ratios of ligation products are
maintained during PCR amplification, since amplification uses only
a single PCR primer. Quantification is relative to one or more
housekeeping genes in the assay.
6
Figure 1. Design of S- and L-Probes
Figure 2. Basic workflow of the DxDirect
method.
For Research Use Only. Not for Use in Diagnostic Procedures
DxDirect Assay Quick Guide
7
For Research Use Only. Not for Use in Diagnostic Procedures
DxDirect Assay Checklist
☐ 1. Add 15 µL DirectReact to each well of the reaction plate.
☐ 2. Add 50 µL of the DxCollect Blood Sample to each well of the reaction plate—MIX WELL.
☐ 3. Add 15 µL of the DirectMix A to each well of the reaction plate.
☐ 4. Add 20 µL of pre-mixed DirectMix B/DirectMix C to each well of the reaction plate—MIX WELL.
☐ 5. Incubate the reaction plate in a PCR Thermal Cycler:
Step
1
2
3
Thermal Cycler Protocol—Ligation
Temperature
Duration
55°C
5 min
80°C
10 min
55°C
45 min
☐ 6. Remove the reaction plate and add 5 µL of pre-mixed M270 Beads to each well of the reaction
plate—MIX WELL.
☐ 7. Incubate the reaction plate in a PCR Thermal Cycler:
Thermal Cycler Protocol—Product Capture
Step
Temperature
Duration
1
55°C
15 min
☐ 8. Place the plate on the magnetic plate for at least 30 seconds.
☐ 9. Remove all supernatant and remove the plate from magnetic plate.
☐ 10. Add 180 µL DirectWash to each well and re-suspend the beads by pipetting.
Repeat
3 times
☐ 11. Prepare the PCR Master Mix (1:1 mix of DirectPrime and DirectTaq).
☐ 12. Remove all supernatant from the plate and add 10 µL of PCR Mastermix to each well of the
reaction plate—cap wells and vortex for 5 to 10 seconds.
☐ 13. Centrifuge the reaction plate and perform PCR:
Thermal Cycler Protocol—PCR
Step
Temperature
Duration
Hot Start
95°C
2 min
Denaturation
95°C
10 sec
3-Step
Annealing
57°C
20 sec
Cycling
Extension
72°C
20 sec
# Cycles
30
N/A
Hold
4°C
∞
☐ 14. Run the final products on CE according to the manufacturer’s instructions.
8
For Research Use Only. Not for Use in Diagnostic Procedures
Detailed Procedure
1. Sample Preparation
DxDirect has been optimized to work with DxCollect RNA Blood Collection Tubes.
Blood samples collected in DxCollect RNA Blood Collection Tubes are stable at room temperature for up
to 7 days prior to processing. If blood samples are to be stored longer than 7 days, then the DxCollect
Tubes should be stored in a -20°C freezer until processing. Prior to using frozen samples, place the tubes
on the bench top for at least 60 minutes to bring samples to room temperature. Once at room
temperature, vortex the tubes gently and proceed with the procedure.
2. Hybridization and Ligation
STOP
Prior to starting the procedure, prepare both DirectMix A and DirectMix B; please
refer to Appendix A and follow the directions.
1. Dispense the appropriate amount of DirectReact reagent into a labeled 1.5 mL microcentrifuge
tube* for the total number of tests to be performed (15 µL per reaction).
2. Dispense the appropriate amount of DirectMix A into a labeled 1.5 mL microcentrifuge tube* for
the total number of tests to be performed (15 µL per reaction).
3. Mix DirectMix B/DirectMix C by combining 15 µL of DirectMix B and 5 µL DirectMix C per
reaction of both reagents into a labeled 1.5 mL microcentrifuge tube*.
* Alternatively, the reagents can be dispensed into 8-well strip microfuge tubes to facilitate
pipetting with a mutli-channel pipette into the 96-well reaction plate.
4. In a 96‐well PCR plate, dispense 15 μL of DirectReact per well into the appropriate number of
wells. (Keep in mind the total number of wells will depend on the number of replicates per
sample). Per well:
a. Add 50 μL of DxCollect stabilized blood sample into the appropriate wells and mix by
pipetting 2–3 times.
b. Add 15 μL DirectMix A and 20 μL DirectMix B/C mixture to all reaction wells.
c. Mix the reactions in each well by pipetting up and down 5 times with a multi-channel
pipette set at 20 µL.
5. Seal the 96-well reaction plate and place the plate in a PCR thermal cycler pre‐programmed to
perform one cycle each of the following 3 incubation steps:
9
For Research Use Only. Not for Use in Diagnostic Procedures
Thermal Cycler Protocol—Ligation
Step
Temperature
Duration
1
55°C
5 min
2
80°C
10 min
3
55°C
2 hr 45 min
3. Target Capture
1. At the end of the 55°C incubation, remove the plate from the thermocycler and after carefully
opening the sealed plate, add 5 µL of the DynaBead Magnetic Beads into each reaction using a
multi-channel pipette. Mix the reactions in each well by pipetting up and down 5 times with a
multi-channel pipette set at 20 µL. Return the reaction plate to the thermal cycler and incubate
for an additional 15 minutes at 55°C.
2. At the end of 15 minutes, remove the reaction plate and place onto a magnetic bead capture
plate for at least 30 seconds. During capture, fill a reagent reservoir suitable for multi-channel
pipetting with DirectWash solution.
3. Remove all ligation reaction solution from the sample plate using an 8-channel 200 µL pipette
set to 180 µL and discard the solution into a biohazard waste container. Avoid pipetting out the
beads.
NOTE: It is important that all of the reaction mixture is removed from the reaction plate. Carry-over
of the reaction mixture may result in subsequent assay failures. Visually inspect the bottom of the
plate to ensure that all liquid is removed from all reaction wells. If any liquid is visible in a well, place
the plate back on the magnetic plate and pipette out the remaining liquid.
4. After removing the reaction solution, visually verify that the beads are on the side of each
reaction well. If beads are not observed in the wells, this indicates loss of the beads/ligation
reaction product, which will result in low signals.
5. Take the reaction plate off the magnetic plate, place the plate on a 96-well plate holder, and add
180 µL of DirectWash into all reaction wells using a multi-channel pipette. Gently re-suspend the
beads by pipetting the solution up and down two times and return the plate to the magnetic
capture plate.
NOTE: Avoid introducing bubbles by gently pipetting the bead mixture during resuspension.
6. Repeat for a total of 3 washes. At the end of the third wash, do NOT remove the wash solution
from the beads.
NOTE: Be sure to visually confirm the magnetic capture of beads before discarding the supernatant
between washes.
4. Amplification
1. Prepare the DirectTaq PCR Master Mix by combining 5 µL of DirectTaq and 5 µL of DirectPrime
for a total volume of 10 µL per reaction.
10
For Research Use Only. Not for Use in Diagnostic Procedures
2. Return the reaction plate to the magnetic capture plate for at least 30 seconds and remove the
final wash solution from the reaction plate. Visually inspect the bottom of the plate to ensure
that all liquid is removed from all wells. If any liquid is visible in a well, place the plate back on
the magnetic plate and aspirate as much remaining wash liquid as possible.
3. Remove the reaction plate from the magnetic capture plate and immediately add 10 µL of the
prepared DirectTaq PCR Master Mix into each well.
NOTE: Dispensing the DirectTaq PCR Master Mix directly onto the magnetic beads makes it easier to
re-suspend the beads with minimal effort.
4. Cap or seal all reaction wells in preparation for PCR. Re-suspend the magnetic beads by
vortexing the plate at a medium-high setting for 5 s to 15 seconds and briefly centrifuge to
pellet the beads. Visually inspect the bottom of the plate to ensure that all beads are immersed
in the PCR Master Mix. If any beads adhere to the sides of the wells exposed to air, vortex the
plate at medium-high setting for 5 seconds and centrifuge the plate at medium speed for 5
seconds.
5. Transfer the plate to a PCR thermal cycler with a hot lid pre-programmed to the following
thermal cycling conditions.
Thermal Cycler Protocol—PCR
Step
Temperature Duration
Hot Start
95°C
2 min
Denaturation
95°C
10 sec
3-Step
Cycling; 30
Annealing
57°C
20 sec
cycles
Extension
72°C
20 sec
Stop
4°C
Hold
6. After thermal cycling is completed, remove the reaction plate and briefly centrifuge. Transfer
the reaction plate to the magnetic capture plate to capture the beads for at least 30 seconds.
5. Detection
1. Prepare sufficient quantities of CE master mix by combining 0.5 µL of the Size Standard and 17.5
µL of Formamide solutions per reaction.
2. Dispense 18 µL of CE Master Mix solution into the appropriate number of wells for the CE
analysis plate.
3. Transfer 2 µL of final PCR reaction product from each reaction well of the reaction plate to the
CE analysis plate while it is still on the magnetic plate. Seal the plate with a plate septum. Vortex
the CE analysis plate at medium speed for 5 seconds and briefly centrifuge.
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For Research Use Only. Not for Use in Diagnostic Procedures
NOTE: Avoid transferring magnetic beads during pipetting. They may interfere with the injection
process during CE.
4. Visually inspect the bottom of the CE plate to ensure that no bubbles are present at the bottom
of any sample well. If any bubbles are observed, re-centrifuge the plate.
5. Run samples by CE according to manufacturer’s user manual. For the ABI-3500 Genetic Analyzer,
the following settings are recommended:
Description
Assay
Polymer type
Capillary Array
Oven temperature
Sample Injection Time
Sizing standard
12
Recommendation
Standard Fragment Analysis
POP-6 or POP-7
50 cm
60°C (default)
30 sec
GeneScan™ 600 LIZ® Size
Standard v2.0
For Research Use Only. Not for Use in Diagnostic Procedures
DxDirect Assay Worksheet: Ligation and Target Capture
Pre-Dispense of DirectReact in Preparation for Addition to Samples
Number of reactions N = _____
Volume per
Volume
Total Volume
Item Description
Reaction, (µL)
Required, (µL)
to Add (µL)
DirectReact
15
Dispense into a 1.5 mL microtube
or 8-microtube strip
15 X 1.2 X N
Total Volume ÷ 8.35
Pre-Dispense of DirectMix A in Preparation for Addition to Samples
Number of reactions N = _____
Volume per
Volume
Total Volume
Item Description
Reaction, (µL)
Required, (µL)
to Add (µL)
DirectMix A
15
Dispense into a 1.5 mL microtube
or 8-microtube strip
15 X 1.2 X N
Total Volume ÷ 8.35
Pre-Dispense of DirectMix B/C in Preparation for Addition to Samples
Number of reactions N = _____
Volume per
Volume
Calculated Volume
Item Description
Reaction, (µL)
Required, (µL)
to Add (µL)
DirectMix B
15
15 X 1.2 X N
DirectMix C
5
5 X 1.2 X N
Total Volume
20
20 X 1.2 X N
Dispense into a 1.5 mL microtube
or 8-microtube strip
Total Volume ÷ 8.35
Pre-Dispense of DirectBeads in Preparation for Addition to Samples
Number of reactions N = _____
Volume per
Volume
Total Volume
Item Description
Reaction, (µL)
Required, (µL)
to Add (µL)
DirectBeads
5
Dispense into a 1.5 mL microtube
or 8-microtube strip
13
5 X 1.2 X N
Total Volume ÷ 8.35
For Research Use Only. Not for Use in Diagnostic Procedures
DxDirect Assay Worksheet: Amplification Setup
Pre-Dispense of PCR Master Mix in Preparation for Addition to Beads
Item Description
Volume per
Reaction, (µL)
Volume
Required, (µL)
DirectTaq
5
5 X N X 1.2 =
DirectPrime
5
5 X N X 1.2 =
Total Volume
10
10 X N X 1.2 =
Dispense into a 1.5 mL microtube
or 8-microtube strip
Calculated Volume
to Add (µL)
Total Volume ÷ 8.35
Preparation of Formamide/Size Standard mixture
Item Description
Volume per
Reaction, (µL)
Volume
Required, (µL)
Formamide
17.5
17.5 X N X 1.2 =
GeneScan 600 LIZ Size
Standard v2.0
0.5
0.5 X N X 1.2 =
Total Volume
18
18 X N X 1.2 =
Dispense into a 1.5 mL microtube
or 8-microtube strip
Calculated Volume
to Add (µL)
Total Volume ÷ 8.35
Reagent Volumes for DxDirect Reactions (includes 20% extra volume)
Volume for
24
reactions
Volume for
48
reactions
Volume for
72
reactions
Volume for
96
reactions
Formulation
Reagents
Volume for
1 reaction
NA
DirectReact
15 µL
432 µL
864 µL
1296 µL
1728 µL
NA
DirectMix A
15 µL
432 µL
864 µL
1296 µL
1728 µL
DirectMix
B/C
DirectMix B
15 µL
432 µL
864 µL
1296 µL
1728 µL
DirectMix C
5 µL
144 µL
288 µL
432 µL
576 µL
NA
M270 Beads
5 µL
144 µL
288 µL
432 µL
576 µL
PCR Master
Mix
DirectTaq
5 µL
144 µL
288 µL
432 µL
576 µL
DirectPrime
5 µL
144 µL
288 µL
432 µL
576 µL
Formamide
17.5 µL
504 µL
1008 µL
1512 µL
2016 µL
LIZ Size
Standard
0.5 µL
14.4 µL
28.8 µL
43.2 µL
57.6 µL
CE Loading
Master Mix
14
For Research Use Only. Not for Use in Diagnostic Procedures
Appendix A
General Guidelines for Preparation of DirectMix A and B
Before performing the DxDirect assay, DirectMix A and B must be prepared beforehand.
DirectMix A and B contain the probes that are used in the assay and are custom designed for your
specific assay. For designing DxDirect probes (L-, S-, and Target Capture (TC)-Probe sets), please refer to
the separate manual on probe design and the use of the AlleleIDTM software (Premiere Biosoft).
DirectMix A contains S-probes in a diluent of 1XTE/1mM DTT.
DirectMix B contains L-probes plus TC-probes in a diluent of 1XTE buffer.
When analyzing DxDirect assays on the ABI CE platforms, it is preferable that gene-specific amplicons
generate peak heights of at least 500 Relative Fluorescent Units (RFUs). Signals below 500 RFU can result
in higher technical variability. Typically, the upper range of signaling for any given fragment should be a
peak height of 10,000 to 15,000 RFU. The preferred upper limit of peak heights will depend on whether
the gene is a normalizer or is expected to be up or down regulated. If signals are over 15,000 RFU, there
is a risk that the peak height will be off-scale (over 28,000 RFU). As a threshold, any signals lower than
100 RFU should be excluded from the analysis as background.
For initial testing, the recommended final (in-assay) concentration is 200 pM for S- and L- probes and
400 pM for TC-probes. If gene-specific signals generated from the assay are too low, then the S-/L-probe
concentration can be increased up to 800 pM. In the case of signals that are too high, the S-/L-probe
concentrations can be lowered to as low as 50 pM. If a further decrease in signal is desired, an S-Probe
Attenuator (SA-) Probe can be used (for further information see “DxDirect: Use of Attenuated S-Probes
to Modulate Signal Magnitude for Specific Genes” Application Note available from DxTerity Technical
Support). The concentrations of matched S- and L-probes should always be the same. In the case of
attenuators, the concentration of an L-probe for any given gene should be equal to the sum of the
concentration of the respective S-probe plus SA-probe.
In addition, it is also possible to modulate all signals in a given sample by changing the CE injection time.
This results in a reduction of raw signals proportional to the change in injection time. For example,
decreasing the CE injection time from 30 seconds to 15 seconds will result in a 50% decrease in the peak
heights of all peaks (including the LIZ internal Size standard).
Quantification of DxDirect Probes
Upon receiving your probe sets, it is recommended that you confirm the concentration for each oligo by
UV absorbance analysis. Measure the absorbance value of the probes at 260 nm and calculate the
concentration of the stock probe concentration using the extinction coefficient value provided with the
oligos.
15
For Research Use Only. Not for Use in Diagnostic Procedures
Creation of Daughter Tubes
Dilute 100 L of 10 µM stock probe with 900 l of diluent to create 1 M working stocks of all probes.
For S-probes, use a diluent of 1X TE/1 mM DTT. For TC- and L-probes, use a diluent of 1X TE.
Formulation Directions for DirectMix A
NOTE: The probes in DirectMix A are at a concentration 6.67-fold higher than in the final mix. Therefore,
if the desired final in-assay concentration of a probe is 100pM, the probe should be formulated in
DirectMix A at a concentration of 667 pM.
1) Determine the volume of DirectMix A required. 15L of DirectMix A is required for a single
reaction. Therefore, 15.0 mL of DirectMix A is sufficient for approximately 1,000 reactions.
2) Calculate the volumes of the 1 M daughter tube S-probes (and SA probes if required) that
should be added to obtain the desired final concentrations. A good starting point is an inreaction concentration of 200 pM S-probe per gene.
3) Add the calculated volume of each probe into an appropriately sized container.
4) After all probes have been added (and before the diluent is added), heat-activate the S-probe
mix in a thermal cycler for 2 minutes at 95˚C.
5) Transfer the S-probes back to the container used for formulation and add the appropriate
volume of diluent (1X TE/1 mM DTT).
Formulation Directions for DirectMix B
1) Determine the volume of DirectMix B required. 15 L of DirectMix B is required for a single
reaction. Therefore, 15.0 mL of DirectMix B is sufficient for approximately 1,000 reactions.
2) Calculate the volumes of the 1 M daughter tube L-probes and TC-probes that should be added
to obtain the desired final concentrations. A good starting point is an in-reaction concentration
of 200 pM L-probe per gene and 400 pM TC-probe per gene).
3) Add the calculated volume of each probe into an appropriately sized container.
4) Add the appropriate volume of diluent (1X TE).
16
For Research Use Only. Not for Use in Diagnostic Procedures
Contact Information
For technical questions regarding the DxDirect Assay and Probe Design, contact:
DxTerity Technical Support
Telephone: 424.772.DXT9
e-mail: [email protected]
For ordering or general questions regarding DxDirect or DxCollect products, contact:
DxTerity Customer Support
Telephone: 424.772.DXT8
e-mail: [email protected]
19500 S Rancho Way, Suite 116
Rancho Dominguez, CA 90220
United States of America
Phone 310.537.7857
Email [email protected]
17
For Research Use Only. Not for Use in Diagnostic Procedures
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