Assay development guidelines and protocol notes
This topic explains the following:
• Each step of the protocol so that you can optimize the Immobilization protocol to your particular experimental design
• Automation movements during the protocol
For details on how to use the Experiments Editor, see Using Experiment IDs.
Protocol stepwise guidelines
Protocol step | Guidelines and notes |
|---|---|
Number of Full Columns of Cartridges | This setting is critical to set the proper force used to mount the cartridges. To obtain expected instrument performance, ensure that the column selection is correct. If the column selection is: • Greater than the actual number of columns used, the Bravo Platform will apply too much force when mounting the cartridges, which can damage both the cartridges and the AssayMAP syringes in the head. For example, if the software specifies 12 columns, but only 1 column of cartridges are in the seating station, the head will apply 12 times more force than what is required. • Less than the actual number of columns used, the Bravo Platform will not apply enough force to seat the cartridges properly. For example, if the software specifies 1 column, but 12 columns of cartridges are in the seating station, the head will apply 1/12th the force required to seat the cartridges properly. In this case, cartridges may fall off during the run or the volume of liquid that moves across the cartridge bed may be variable due to liquid moving past the syringe cartridges seal into the cartridge cup. Default: 1 Range: 1-12 |
Initial Syringe Wash | This step flushes any potential contaminants from the syringes at the wash station before the cartridges are mounted. During each Initial Syringe Wash cycle, the head aspirates 250 µL into the syringes from the wash station chimneys and then moves by a fixed offset between the chimneys to dispense to waste. This step is selected by default. Wash Cycles. Increasing the number of wash cycles may clean the syringes better. However, more cycles increases the total run time and causes wear on the syringes. • Default: 3 • Practical: 3–5 • Range: 0–10 |
Prime | This step removes entrained air from the packed resin bed and properly wets the surface of the resin. In preparation for priming, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Priming Buffer is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. The Prime step aspirates the Priming Buffer into the syringes, mounts the cartridges, and then dispenses the buffer through the cartridges into the wash station. The cartridges are parked at the seating station and the syringes are washed at the wash station. The AssayMAP affinity purification cartridges (PA-W, SA‑W, and PG‑W) typically used with this application contain affinity ligands (proteins) covalently immobilized onto resin. These cartridge types should be primed with aqueous buffers containing no or low amounts of organic solvent or protein denaturants. Because the Equilibration Buffer is drawn from the same reservoir as the Priming Buffer, buffers that favor analyte binding with minimal non-specific binding should be used for both priming and equilibration. This step is selected by default. Volume (µL). The default volume should be sufficient to wet and remove entrained air from the resin bed. Using less than the default volume may leave air in the resin bed. Using more than the default volume is unnecessary and increases run time. • Volume for 5 µL cartridges: – Default: 100 – Practical: 100–250 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Prime tasks except syringe washing. Flow rate (µL/min). A flow rate slower than the default value diminishes the ability to effectively remove entrained air from the resin bed. A flow rate faster than the default is not required and has not been tested. • Default: 300 • Practical: 300 • Range: 0.5–500 Wash cycles. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 1 • Practical: 1–3 • Range: 0–10 |
Equilibrate | This step ensures that the resin bed is fully equilibrated with a solution that provides the optimal chemical conditions for binding during the Load Samples step. In preparation for equilibration, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Equilibration Buffer is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. During the Equilibrate step, the Equilibration Buffer is aspirated into the syringes, the cartridges are mounted, and then the buffer is dispensed through the cartridges into the wash station. The cartridges are parked at the seating station and the syringes are washed at the wash station. This step is selected by default. Volume (µL). The default volume is equal to 10 column volumes, which should be sufficient for complete buffer exchange. Using less than the default volume may not fully equilibrate the resin bed. Using more than the default volume is unnecessary and increases run time. • Volume for 5 µL cartridges: – Default: 50 – Practical: 50–100 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Equilibrate step tasks except syringe washing. Flow rate (µL/min). A flow rate slower than the default rate will likely have no benefit, but will increase the total assay time. A flow rate faster than 20 µL/min may not equilibrate through the pores in the beads. • Default: 10 • Practical: 5–20 • Range: 0.5–500 Wash cycles. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 1 • Practical: 1–3 • Range: 0–10 |
Load Samples | This step allows the target analytes to bind to the surface chemistry of the resin bed. No liquid is removed or added to the cartridge cups before the sample loading begins. The assumption is that there is still liquid in the cups from the equilibration step that will prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. This step aspirates sample into the syringes, and then performs an external syringe wash at the wash station to remove any sample remaining on the outside of the probes before mounting the cartridges. The samples are dispensed through the cartridges into the Flow Through Collection plate or wash station. The exterior of the cartridge tips are washed at the wash station to remove sample on the exterior of the cartridges, the cartridges are parked at the seating station, and the syringes are washed at the wash station. The protocol accommodates sample volumes up to 1000 µL to be dispensed through the cartridges. Although, the form permits you to enter smaller volumes, the minimum advisable sample volume to be loaded onto an AssayMAP cartridge is 10 µL. Each syringe has a maximum capacity of 250 µL. When sample volumes are greater than 250 µL, the protocol will iteratively load samples onto cartridges. To determine the number and volume of the iterative load steps, the protocol uses the following formulas: • # of times to load = total sample volume/250, where # times to load is rounded up to nearest integer • volume of each load = sample volume/# of times to load For example, if the total sample volume is 900 µL, then: # times to load = 900/250 = 3.6, which is rounded up to 4 volume of each load = 900/4 = 225 If Collect Flow Through is selected for the Load Samples step, be sure that the Flow Through Collection plate has sufficient maximum well capacity. For details, see the Labware Reference Guide in the Literature Library page of the Protein Sample Prep Workbench.  Be sure to include the recommended labware-dependent volume overage to prevent air from entering the cartridge. For more information, see Preparing the sample plates.To determine the volume of sample to load, see Determining the volume of sample to load. This step is selected by default. |
Volume (µL): The volume of sample to load should be balanced with the sample concentration and the mass capacity of the cartridge. The lower the sample volume, the higher the percentage of the total volume is overage. To minimize sample loss, Agilent recommends diluting small volume samples. Large sample volumes (> 250 µL) may require slightly more excess sample due to evaporation. • Default: 100 • Practical: 10–1000 • Range: 0–1000 Note: Setting the volume to zero skips all Load Samples tasks except syringe washing. Flow rate (µL/min). The optimum sample loading flow rate requires balancing the speed of the assay and desired recovery. When setting the flow rate, be aware that the quantitative binding capacity is inversely proportional to the flow rate. Therefore, the maximum possible quantitative binding capacity is only obtained with very slow sample loading flow rates. If the amount of sample that you want to capture is significantly lower than the total possible qualitative binding capacity, you will be able to use a faster flow rate while maintaining quantitative binding. Using flow rates slower than 5 µL/min may not significantly increase analyte binding, but this is highly dependent on the molar ratio of the capture ligand compared to the target molecule. For examples of cases with flow rates of less than 5 µL/min, see Agilent app notes 5991-9010EN and 5991-8445EN in the Reference library. • Default: 5 • Practical: – 2–10 (5 µL cartridges) – 5–20 (25 µL cartridges) • Range: 0.1–500 Wash cycles. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 3 • Practical: 2–5 • Range: 0–10 | |
Collect Flow Through | If this step is selected, the sample flow-through from the Load Samples step is dispensed in the Flow Through Collection plate. If this step is not selected, the flow-through from the Load Samples step is dispensed directly into the wash station. The Collect Flow Through step is skipped if the Load Samples step is not conducted. This step is selected by default. |
Cup Wash 1 | This step removes the residual sample liquid that may remain above the resin bed after the Load Samples step. The Cup Wash 1 step aspirates Cartridge Wash Buffer 1 into the syringes and then dispenses it into the cups of the parked cartridges. This liquid plus any residual liquid from samples is aspirated from the cartridge cups. The protocol ensures that no cartridges are stuck to the probes before dispensing the liquid into the wash station, and then washing the syringes at the wash station. This step is selected by default. Volume (µL). Using a volume less than the default may be insufficient for cup washing, while using a volume >50 µL may offer little benefit. • Default: 25 • Practical: 25–50 • Range: 0–100 Note: Setting the volume to zero skips all Cup Wash tasks. Wash cycle. Each cycle comprises one cup wash and one syringe wash. • Default: 3 • Practical: 3–5 • Range: 0–10 |
Internal Cartridge Wash 1 | This step uses Cartridge Wash Buffer 1 to wash non-specifically bound molecules from the resin bed. In preparation for Internal Cartridge Wash 1, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Cartridge Wash Buffer 1 is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. For the wash operation, this step aspirates Cartridge Wash Buffer 1 into the syringes, mounts the cartridges, and then dispenses the buffer through the cartridges into the Flow Through Collection plate or wash station. The exterior of the cartridge tips are washed at the wash station to remove any remaining buffer on the cartridge exterior, the cartridges are parked at the seating station, and the syringes are washed at the wash station. If the Load Samples step is selected, the first 5 µL (5 µL cartridges) or 25 µL (25 µL cartridges) of Cartridge Wash Buffer 1 is dispensed as a sample chase at the Load Samples flow rate. Next, the Internal Cartridge Wash volume minus the chase volume is dispensed at the Internal Cartridge Wash flow rate. The sample chase ensures that the sample volume in the cartridges at the end of the sample load moves through the cartridge bed at the same rate as the rest of the sample. This step is selected by default. Volume (µL). Volumes higher than the default volume (10 column volumes) may improve the purification marginally but also increases the run time. Volumes lower than the default volume may be insufficient for efficient cartridge washing. • Volume for 5 µL cartridges: – Default: 50 – Practical: 50–100 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Internal Cartridge Wash tasks except syringe washing. Flow rate (µL/min). A rate slower than the default flow rate will have little benefit, but will increase the total assay time. A rate faster than 20 µL/min may not equilibrate through the pores in the beads, resulting in incomplete washing. • Default: 10 • Practical: 5–20 • Range: 0.5–500 Wash cycle. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 3 • Practical: 2–5 • Range: 0–10 |
Collect Flow Through | If this step is selected, the flow-through from Internal Cartridge Wash 1 is dispensed in the Flow Through Collection plate. If the Collect Flow Through step is not selected, the flow-through from Internal Cartridge Wash 1 is dispensed at the wash station. This step is not selected by default. |
Load Blocking Reagent | This step allows a reagent of defined composition to be flowed through the cartridge after ligand immobilization to help minimize non-specific binding in subsequent target purification steps. The Load Blocking Reagent step may be especially helpful when a sub-saturating amount of ligand is bound to the resin in the cartridge. In preparation for blocking, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Blocking Reagent is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. The Load Blocking Reagent step aspirates Blocking Reagent into the syringes, performs an external syringe wash at the wash station, mounts the cartridges, dispenses the Blocking Reagent through the cartridges at the specified flow rate to either the Flow Through Collection plate or the wash station. An external cartridge wash is performed at the wash station to remove any blocking reagent on the outside of the cartridge. The cartridges are parked at the seating station and the syringes are washed at the wash station. Select the Load Blocking Reagent step to minimize to non-specific binding during subsequent target purification steps. This step is selected by default. Volume (µL). The volume of Blocking Reagent to aspirate into the syringes and dispense through the cartridges. • Volume for 5 µL cartridges: – Default: 50 – Practical: 50–100 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Load Blocking Reagent tasks except syringe washing. Flow rate (µL/min). A rate slower than the default will likely have no benefit, but will increase the total assay time. A rate faster than 20 µL/min may not equilibrate through the pores in the beads, which may result in incomplete blocking. • Default: 5 • Practical: – 2–10 (5 µL cartridges) – 5–20 (25 µL cartridges) • Range: 0.1–500 Wash cycles. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 3 • Practical: 2–5 • Range: 0–10 |
Collect Flow Through | If this step is selected, the liquid eluted during the Load Blocking Reagent step is dispensed into the Flow Through Collection plate. If this step is not selected, the flow-through from the Load Blocking Reagent step is dispensed into the wash station. Select this step if you want to collect the Blocking Reagent flow-through. This step is not selected by default. |
Cup Wash 2 | This step removes the residual buffer that may remain above the resin bed after the Load Blocking Reagent step. This step aspirates Cartridge Wash Buffer 2 and then dispenses it into the cups of the parked cartridges. This liquid plus any residual liquid from the previous step is aspirated from the cartridge cups. Any cartridges that stuck to the probes during the cup wash are removed at the seating station, and then the liquid in the syringes is dispensed into the wash station. The syringes are washed at the wash station. This step is selected by default. Volume (µL). A volume less than the default may be insufficient for cup washing, while a volume >50 µL may offer little benefit. • Default: 25 • Practical: 25–50 • Range: 0–100 Note: Setting the volume to zero skips all Cup Wash 2 tasks. Wash cycle. Each cycle comprises one cup wash and one syringe wash. • Default: 3 • Practical: 3–5 • Range: 0–10 |
Internal Cartridge Wash 2 | This step uses Cartridge Wash Buffer 2 to wash non-specifically bound molecules and Blocking Reagent from the resin bed. In preparation for Internal Cartridge Wash 2, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Cartridge Wash Buffer 2 is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. For the wash operation, this step aspirates Cartridge Wash Buffer 2 into the syringes, mounts the cartridges, and then dispenses the buffer through the cartridges at the specified flow rate into the Flow Through Collection plate or wash station. The exterior of the cartridge tips are washed at the wash station to remove any remaining buffer from the previous step on the cartridge exterior, the cartridges are parked at the seating station, and the syringes are washed at the wash station. This step is selected by default. Volume (µL). Volumes higher than the default volume (10 column volumes) may improve the purification marginally but will also increase the run time. Volumes lower than the default volume may be insufficient for efficient cartridge washing. • Volume for 5 µL cartridges: – Default: 50 – Practical: 50–100 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Internal Cartridge Wash tasks except syringe washing. Flow rate (µL/min). A rate slower than the default flow rate will have little benefit, but will increase the total assay time. A rate faster than 20 µL/min may not equilibrate through the pores in the beads, resulting in incomplete washing. • Default: 10 • Practical: 5–20 • Range: 0.5–500 Wash cycle. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 3 • Practical: 2–5 • Range: 0–10 |
Collect Flow Through | If this step is selected, the liquid eluted during Internal Cartridge Wash 2 is dispensed into the Flow Through Collection plate. If the Collect Flow Through step is not selected, the flow-through is dispensed into the wash station. Select this step if you want to collect the flow-through generated during Internal Cartridge Wash 2. This step is not selected by default. |
Stringent Syringe Wash | This step cleans the syringes with the Stringent Syringe Wash Buffer. The Stringent Syringe Wash step aspirates the Stringent Syringe Wash Buffer into the syringes, draws the buffer through a full syringe stroke to ensure the entire syringe is rinsed, and then dispenses the buffer into the wash station. The syringes are then washed at the wash station. This step is selected by default. Volume (µL). Volumes higher than the default volume are unlikely to improve the syringe cleaning but will increase the run time. Volumes lower than the default volume may be insufficient for efficient syringe washing. • Default: 50 • Practical: 50–100 • Range: 0–250 Note: Setting the volume to zero skips all Stringent Syringe Wash tasks. Wash cycle. A wash cycle is a stringent syringe wash followed by a basic syringe wash at the wash station. • Default: 2 • Practical: 2–5 • Range: 0–10 |
Re-Equilibrate | This step conditions the resin and immobilized ligand to prepare them for the next step in the workflow, for example, running the Affinity Purification protocol. In preparation for re-equilibration, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station, 10 µL of Equilibration Buffer is aspirated into the syringes and then dispensed into the cartridge cups to prevent potential air gaps from being introduced when the cartridges are seated on the syringe probes. The Re-Equilibration step aspirates the Equilibration Buffer into the syringes, mounts the cartridges, and then dispenses the buffer through the cartridges into the wash station. The cartridges are parked at the seating station and the syringes are washed at the wash station. Do not allow cartridges to dry out. After re-equilibration, use immediately for affinity purification or store them short term in the receiver plate containing 200 µL of Equilibration Buffer per well. This step is selected by default. Volume (µL). The default volume is equal to 10 column volumes, which should be sufficient for complete buffer exchange. Using less than the default volume may not fully re-equilibrate the resin bed. Using more than the default volume is unnecessary and increases run time. • Volume for 5 µL cartridges: – Default: 50 – Practical: 50–100 – Range: 0–250 • Volume for 25 µL cartridges: – Default: 250 – Practical: 250 – Range: 0–250 Note: Setting the volume to zero skips all Re-Equilibrate tasks except syringe washing. Flow rate (µL/min). A flow rate slower than the default rate will likely have no benefit, but will increase the total assay time. A flow rate faster than 20 µL/min may not equilibrate through the pores in the beads in the cartridge resin bed. • Default: 10 • Practical: 5–20 • Range: 0.5–500 Wash cycles. The number of syringe wash cycles to perform at the end of this step. 250 µL of DI water is used for each syringe wash cycle. • Default: 1 • Practical: 1–3 • Range: 0–10 |
Final Syringe Wash | This step uses the wash station to flush potential contaminants from the syringes. Before the final syringe wash begins, 20 µL of air is aspirated into the syringes, the probes go into the cartridge cups to a depth that is just short of the normal engagement position, liquid in the cups is removed by a 60 µL aspiration and then discarded into the wash station. No solution is added into the cartridge cups. Note: If the Final Syringe Wash is skipped, the 10 µL of elution buffer will remain in the cartridge cups. During each Final Syringe Wash cycle, the head aspirates 250 µL into the syringes from the wash station chimneys, and then moves by a fixed offset between the chimneys to dispense the syringe contents to waste. Note: In cases where carryover is a major concern, increasing the number of wash cycles may provide improved washout, but with a cost of increased assay time and reduced syringe lifetime. The best practice is to use the Syringe Wash utility to wash the syringes between runs with stringent wash solutions. This step is selected by default. Wash Cycles: • Default: 3 • Practical: 3–5 • Range: 0–10 |
Automation movements during the protocol
This section describes the basic movements of the AssayMAP Bravo Platform during the Immobilization protocol using the default method. Changing the selections or parameters will alter the movements.
Protocol step | Head moves to deck location... | Action |
|---|---|---|
Starting protocol | 2 | Parks any cartridges that may have been mounted on the head from a protocol that had been previously aborted. |
1 | Dispenses any liquid remaining in the syringes into the wash station. | |
Initial Syringe Wash | 1 | Washes the syringes the specified number of times. |
Prime | 2 | Aspirates 20 µL of air above this location, moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys, and then washes the exterior of the syringe probes. | |
3 | Aspirates 10 µL of Priming & Equilibration Buffer for the cartridge air-gap-prevention step. | |
2 | Dispenses the 10 µL of buffer into the cartridge cups and exercises the cartridges off task. | |
3 | Aspirates the Priming Buffer. | |
2 | Mounts the cartridges on the head. | |
1 | Dispenses the Priming & Equilibration Buffer through the cartridges into the wash station, and then washes the exterior of the cartridge tips. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes. | |
Equilibrate | 2 | Aspirates 20 µL of air above this location, moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys, and then washes the exterior of the syringe probes. | |
3 | Aspirates 10 µL of Priming & Equilibration Buffer for the cartridge air-gap-prevention step. | |
2 | Dispenses the 10 µL of buffer into the cartridge cups and exercises the cartridges off task. | |
3 | Aspirates the Priming & Equilibration Buffer. | |
2 | Mounts the cartridges on the head. | |
1 | Dispenses the buffer through the cartridges to the wash station. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes. | |
Load Samples | 4 | Aspirates samples into the syringes. |
1 | Washes the exterior of the syringe probes. | |
2 | Mounts the cartridges on the head. | |
7 | Dispenses sample through the cartridges to load sample. Collects flow-through in the Flow Through Collection plate. | |
1 | Washes the cartridge exteriors at the wash station. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes. | |
Cup Wash 1 | 5 | Aspirates Cartridge Wash Buffer 1 into the syringes. |
2 | Performs the cup wash and exercise the cartridges off task. | |
1 | Dispenses buffer into the wash station. | |
1 | Washes the syringes. | |
Internal Cartridge Wash 1 | 2 | Aspirates 20 µL of air above this location, moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys. | |
5 | Aspirates 10 µL of Cartridge Wash Buffer for the cartridge air-gap prevention step. | |
2 | Dispenses the 10 µL of buffer into the cartridge cups and exercises the cartridges off task. | |
5 | Aspirates Cartridge Wash Buffer 1 into the syringes. | |
2 | Mounts the cartridges on the head. | |
1 | Dispenses 5 µL (5 µL cartridges) or 25 µL (25 µL cartridges) Cartridge Wash Buffer 1 through the cartridges at the Load Samples flow rate for the sample chase step. | |
1 | Dispenses the remaining Cartridge Wash Buffer 1 through the cartridges at the Internal Cartridge Wash 1 flow rate. | |
1 | Washes the exterior of the cartridge tips. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes at the wash station. | |
Load Blocking Reagent | 9 | Aspirates Blocking Reagent into syringes. |
1 | Washes exterior of probes at the wash station. | |
2 | Mounts the cartridges on the head. | |
1 | Dispenses Blocking Reagent through the cartridges. | |
1 | Washes the cartridge exteriors at the wash station. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes at the wash station. | |
Cup Wash 2 | 6 | Aspirates Cartridge Wash Buffer 2 into the syringes. |
2 | Performs the cup wash and exercise the cartridges off task. | |
1 | Dispenses buffer into the wash station. | |
1 | Washes the syringes at the wash station. | |
Internal Cartridge Wash 2 | 2 | Aspirates 20 µL of air above this location, moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys, and then washes the exterior of the syringe probes. | |
6 | Aspirates 10 µL of Cartridge Wash Buffer 2 for the cartridge air-gap prevention step. | |
2 | Dispenses the 10 µL of buffer into the cartridge cups and exercises the cartridges off task. | |
6 | Aspirates Cartridge Wash Buffer 2 into the syringes. | |
2 | Mounts the cartridges on the head. | |
1 | Dispenses Cartridge Wash Buffer 2 through the cartridges. | |
1 | Washes the exterior of the cartridge tips. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes at the wash station. | |
Stringent Syringe Wash | 8 | Aspirates the Stringent Syringe Wash Buffer. |
1 | Dispenses the buffer into the wash station. | |
1 | Washes the syringes at the wash station. | |
Re-Equilibrate | 2 | Aspirates 20 µL of air above this location, moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys, and then washes the exterior of the syringe probes. | |
3 | Aspirates 10 µL of Priming & Equilibration Buffer for the cartridge air-gap-prevention step. | |
2 | Dispenses the 10 µL of buffer into the cartridge cups and exercises the cartridges off task. | |
3 | Aspirate the Priming & Equilibration Buffer. | |
2 | Mounts the cartridges. | |
1 | Dispenses buffer through the cartridges into the wash station. | |
1 | Washes the exterior of the cartridge tips. | |
2 | Parks the cartridges in the seating station. | |
1 | Washes the syringes at the wash station. | |
Final Syringe Wash | 2 | Moves down to just above the cartridge engagement point and aspirates 60 µL, and then exercises the cartridges off task. |
1 | Dispenses into the wash station between the chimneys. | |
1 | Washes the syringes at the wash station. |