December, 2014
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Genotyping Lab Mantis Pipetting Robot Protocol
Overview
The Mantis is a programmable single channel pipetting robot which can pipette
into 96, 384, or 1536 well plates. It is best for small volumes, but can work with
large volumes too. It can pipet into single wells or any number of wells. The
dispensing head uses disposable pipette tips as the reagent reservoirs. It can
transfer a minimum of 0.1 ul to dry wells with REUSABLE LOW VOLUME chips
or 1.0 ul to dry wells with REUSABLE HIGH VOLUME chips. The Mantis greatly
decreases waste (dead) volumes down to about 100 ul. Up to 6 different reagents
can be dispensed in 1 pipetting script.
Precautions
• Only use plates and labware that Paul has entered into the plate type library
and trained. See Paul to add new labware.
• Do NOT put USB drives, floppy disks, or burned CD/DVDs into the robot
computer. We have NO virus software on this computer.
• Do NOT install ANY software on this computer.
• If any genomic DNA is run through any chip or hose, those chips and hoses
MUST be cleaned with bleach and then water! If not, DNA WILL contaminate
every reaction that follows!
• We can use the Mantis and the Biomek robots at the same time by switching to
the appropriate software. Do NOT quit or close any software while either robot is
operating.
• If the software does not find the Mantis, disconnect the USB cable from the
back of the Mantis then reconnect it and try again.
• The Mantis dispenses very uniformly from well to well, but may dispense 3%
to 9% more volume than requested. For precise work, do a gravimetric test on
each chip that you will use.
• Pipetting small volumes will only take 2 minutes per 384 well plate. Volumes
larger than 3 ul will take significantly more time. For example, 200 ul in a 96
deepwell plate may take 17 minutes.
• Always check that you have enough reagent volume in your pipette tips before
pipetting, the robot does NOT check if you have sufficient volume for a dispense.
• The user manual PDF for the Mantis is on the computer desktop.
Operating the Mantis
Prepare the Robot and Dispense List
1. Turn on power. The switch is on the right rear side.
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2. If the wash reservoir is less than half full, fill it with autoclaved NanoPure
water.
3. If the waste reservoir is more than half full, empty it.
4. Mount your plate into an adapter if needed and place it on the Mantis
within the 4 mounting pins. 384 well and 96 well PCR plates REQUIRE
adapter plates.
5. If you will be using the “Water-Hose” position, make sure you have
enough autoclaved NanoPure water in the 50 ml conical. Do NOT touch
the hose with your hands. Wear NEW gloves when adding water and
handling the tubing. Remember, this water will end up in your reactions!
6. Open the Mantis software from the desktop icon. Open a “dispense list”
from the file menu.
7. Dispense lists are grouped by plate type in the “DispenseList” folder. Start
with one from the “Templates” folder. Make a folder with your name in
the “DispenseList” folder. Save your lists to your folder ONLY.
8. Reagents are assigned to chip positions on the Mantis. Do NOT change
them. You can add reagents to your list, remove reagents from your list,
and change the volumes using the “List Designer” on the center and right
side panels of the software. If your reagent is not one of the pre-assigned
reagents, use the “Other” chip position or the “Other-LV” position. You
may also edit complex lists using Excel (press the Excel button in the list
designer). After editing a list in Excel, save the list and quit Excel. The file
will be imported in the Mantis software.
9. Select wells and then enter the volume and press the “enter” key to
change the volumes of all or any of the wells. Remember, the HV chips
can deliver volumes ONLY in increments of 1 ul. The LV chips can deliver
volumes ONLY in increments of 0.1 ul.
10. If you are using the “Water-Hose” position, right-click on the “WaterHose” title in the lower left panel and choose “Attach input to hand”. This
will mount the chip. Then press and hold on the “Manual priming input”
until water drips are seen ejecting continuously from the chip into the
waste. This removes air from the hose and primes the chip. All other
positions are primed just before they are used.
11. Make certain that the hose from the "Water-Hose" reservoir is not in the
path of the pipetting head or it will become tangled.
Dispense
1. Pipette your reagents using a 1,000 ul tip (or smaller). If you click on the
reagent in the center list, you can see the volume required for your
samples. Ensure that you have enough volume for your samples plus 110
ul additional for priming and waste.
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2. Ensure that there is NO air at the very end of the pipette tip before
mounting.
3. Mount the tip onto the correct chip position by inserting the filled pipette
tip into the green chip opening (LV chips have a black opening), hold the
top of the pipette tip firmly and eject the tip from the pipetter. Then press
the tip down with light pressure and rotate the tip half of a turn to seat the
tip.
4. Once all needed reagents have been loaded onto chips, press the green run
arrow to start the dispense.
5. KEEP YOUR HANDS AWAY FROM THE ROBOT WHILE IT IS
PIPETTING.
6. If you need to abort the dispense, press the red “Stop Dispense” button
and wait for the robot to stop.
Cleanup
1. Each chip used MUST be properly washed and dried or reagent will clog
and contaminate the chip and the chip will need to be replaced.
2. Remove pipette tip from a chip by: holding the chip down with one finger,
placing the tip of another finger on the top of the pipette tip to block
reagent drips, then pulling the tip up from the chip.
3. Right-click on your used reagent-chip in the Device Control panel and
select “Attach Input to Hand” to mount a used chip.
4. Click on the “Recover Input” button to force all remaining reagent out of
the chip and back into the pipette tip.
5. Click on the “Wash Input” button to wash the chip. Do this TWICE!
6. Click on the “Recover Input” button to force air into the chip to dry it. Do
this TWICE!
7. Right-click on your used reagent-chip in the Device Control panel and
select “Detach Input from Hand” to put away the chip.
8. Save your dispense list to your folder if needed.
9. Exit the Mantis software and switch off the Mantis robot. Do not
disconnect the USB cable.
10. Return adapters to the Mantis box and clean up all of your used tips.
11. Use KimWipes to dry any liquid from the chips and chip positions,
especially the chip nozzle.
DNA Cleanup
1. If you have run ANY AMOUNT OF GENOMIC DNA through any chip
positions, you MUST BLEACH WASH THAT CHIP! Since the chips are
reusable, we MUST keep them free of template DNA. That is true for the
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Water-Hose position also. PCR primers in a master mix will be washed
out of the chips with a standard water cleaning.
2. Make a 10% bleach solution (100 ul bleach and 900 ul NanoPure water).
3. Fill a 1,000 ul tip with the 10% bleach solution and mount the tip on the
contaminated chip.
4. Right-click on your contaminated chip in the Device Control panel and
select “Attach Input to Hand” to mount the chip.
5. Press and hold on the “Manual priming input” until water drips are seen
ejecting continuously from the chip into the waste tube. Keep priming
until the bleach in the pipette tip is about half used up.
6. Allow the bleach to remain in the chip for 5 to 10 minutes. It does take a
minimum of 5 minutes to destroy the DNA.
7. Press and hold on the “Manual priming input” until all of the bleach in
the pipette tip is gone and only air is exiting the chip.
8. Right-click on your bleached chip in the Device Control panel and select
“Detach Input from Hand” to put away the chip.
9. Fill a NEW 1,000 ul tip with NanoPure water and mount the tip on the
bleached chip.
10. Right-click on the bleached chip in the Device Control panel and select
“Attach Input to Hand” to mount the chip.
11. Press and hold on the “Manual priming input” until water drips are seen
ejecting continuously from the bleached chip into the waste tube. Keep
priming until the water in the pipette tip is about half used up.
12. Allow the water to remain in the chip for 1 to 5 minutes.
13. Press and hold on the “Manual priming input” until all of the water in the
pipette tip is gone and only air is exiting the chip.
14. Click on the “Recover Input” button to force air into the chip to dry it. Do
this TWICE!
15. Right-click on your washed chip in the Device Control panel and select
“Detach Input from Hand” to put away the chip.
16. Save your dispense list to your folder if needed.
17. Exit the Mantis software and switch off the Mantis robot. Do not
disconnect the USB cable.
18. Return adapters to the Mantis box and clean up all of your used tips.
19. Use KimWipes to dry any liquid from the chips and chip positions,
especially the chip nozzle.
Maintenance (Paul Only)
• Backup files every 2 months.
• Add new labware definitions as needed.
• Soak all chips in 10% bleach and run bleach and water through the chips as
needed.
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Mantis Components
Mantis User's Guide
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M ANTIS FRONT
Mantis
Front
Chip Changers
Hand
Chip Changer
Plat e Clamps
Using the chip changers, the Mantis can automatically
dispense up
Arm
to six reagents without user intervention. The chip changers are
compatible with High Volume (HV), Low Volume (LV), High
Volume PFE, Low Volume PFE, Low Volume Continuous Flow (CF),
or High Volume Continuous Flow (CF) chips.
Plate Clamp
Located on the lower-right of the plate holder, the plate clamps
align and secure the plate during the dispensing process.
Plate Holder
Tube Holders
Plat e Holder
The plate holder is compatible with almost all SBS plate types and
deep well blocks.
Tube Holder
The tube holders are compatible with 50 mL Falcon tubes that are
ideal for large volume dispenses or frequently used reagents.
Mantis Back
A TTACHING THE A CCESSORIES Arms
M ANTIS BACK
The Mantis's arms control the hand movements when dispensing,
loading and unloading chips.
We provide the waste and wash accessories that you need to place in Mantis.
Locate the plastic tubular waste andHand
wash stations that were in The
yourhand
accessory
holds the High Volume (HV), Low Volume (LV), High
kit.
Volume PFE, Low Volume PFE, Low Volume Continuous Flow (CF),
or High Volume Continuous Flow (CF) chip during general
● Place the wash stations in the wash station holders on operations.
the back of the
Mantis, and the waste station in the waste station holder.
● Remove the packing tape that holds the wash tube to the Mantis’s body
Waste Station
and put the wash tube in the wash station.
Wash Station 2
Wash Station 1
9
A SSEM BLING THE M ANTIS CHIPS
USB Ports
The Mantis’s chips and chip pallets are packaged separately, but they are
both found in the accessory kit.
Power Switch
Power Port
High Volume/ Low Volume/ High Volume PFE/ Low Volume PFE
Locate the magnetic chip pallet and the High Volume (HV), Low Volume
(LV), High Volume PFE, and/ or Low Volume PFE chips, and snap the chip into a
magnetic chip pallet. Repeat this process for all six chips. Then, put the chip and
chip pallet into the chip changer.
Wast e St at ion
The waste station is a station for the Mantis to deposit waste
liquids.
Mantis Chips
Mantis Reagent Positions
Tube Input Adapter
Wash St at ions
PFE Chip
HV Chip
The wash stations are where the Mantis holds wash solutions and
water, used during the chip washing process. Wash solutions
should go in Wash Station 1, and water should go in Wash
Station 2.
LV Chip
10
Magnetic Chip Pallet
4
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