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FAQ - Microtiter plate format
Which side of the MTP plate is the coated?
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The MTP plates with the following surfaces: Epoxy (E), Aminosilane (AStar & A+), or Aldehyde (AL) are coated on both sides of the glass. The correct side of the MTP plate for spotting is the same side as the 96-well black patterning. The MTP plates that are coated with the 3-D thin film polymer (P) are only coated on one side (the side with the black 96-well patterning).
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Chamfer corner in the patterning indication well A1
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Which is the recommended spotting solution for the MTP-96 plates?
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If the print run is likely to take several hours, then low evaporation print buffers are recommended.
However the best spotting solution to use depends on the coating of the MTP:
- For MTP A+ and AStar Nexterion® Spot A HD, or 50% DMSO is recommended.
- MTP E works well with 3x SSC with 0.75 – 1.5 M betaine or Nexterion® Spot LE.
- MTP P Nexterion® Spot PB, or DMSO containing buffers for peptide arrays are recommended.
More detailed recommendations on suitable spotting, blocking and hybridization buffers are available in the individual product protocols.
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What difficulties are caused by printing the arrays too close to the black PTFE well patterning?
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The well edges in the PTFE patterning have a maximum deviation of ± 100 µm. The printer software should be set up to print the arrays at least 550 µm away from the “theoretical” well edge. If the pins are set to print any closer there is a risk of printing over the patterning. Printing too close to the well edges may also cause problems during the hybridization / incubation step. For example, if the silicone superstructure is not accurately placed, it is possible that some spots may be very close to, or even under the superstructure. In this case it will be impossible to hybridizing or scan these spots.
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Carefully remove the opaque protective backing to expose the adhesive, and apply the superstructure to the plate by holding it at the 4 corners. Once the superstructure is aligned on the plate, press down on each individual well for 3 to 5 seconds to ensure the proper adhesion to the patterned glass.
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Method 2: Without using plate holder
Lay the superstructure upside down on clean flat surface (i.e. so the adhesive layer is uppermost). Then remove the opaque protective backing from the superstructure to expose the adhesive. Turn the printed plate upside down, and carefully place it onto the superstructure.
Important: Make sure that the superstructure is not misaligned with the wells on the glass plate, otherwise it will not fit back into the MTP plate holder. Please press the superstructure around each individual well for 3 to 5 seconds to ensure proper adhesion. Fix the glass plate and attached superstructure into the MTP plate holder.
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How can I avoid cross contamination between neighboring wells on the MTP plate?
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The black patterning is hydrophobic and creates 96 square, 7 by 7 mm wells. The combination of the hydrophobic pattering and the self-adhesive silicone superstructure prevents cross contamination between the wells. It is necessary to put the superstructure accurately on the plate patterning, and press down firmly around each individual well to create a good seal. Inaccurately applying the 96-well superstructure could result in cross contamination, and also risks introducing adhesive into the printed area:
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Cross contamination between wells as a result of an inaccurately applied superstructure
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Tecan Hydroflex™
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SCHOTT’s R&D team has successfully used the Tecan Hydroflex™ with the Nexterion MTP-96 plate.
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The best method to remove the superstructure from the plate is to grab the superstructure with tweezers at one corner and gently pull it up and off. During removal take care that all the adhesive is removed from the plate. Also see the question below.
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Is there an optimal temperature for removing the superstructure from the MTP plate?
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The optimal temperature for successfully removing the superstructure from the MTP patterned glass plate is between 31 and 41°C.
At lower temperatures (20 to 30°C), the double-sided adhesive foil attached to the silicone superstructure can delaminate, resulting in two components.
At temperatures above 41°C the adhesive becomes very tacky, and there is a risk of filaments being created during the removal of the superstructure. These filaments might contaminate the arrays and adversely affect the subsequent image analysis.
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How can I avoid evaporation of the hybridization solution during the hybridization?
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To avoid evaporation of the hybridization solution, it is important that the clear sealing sheet supplied with the 96-well superstructure is used to seal the top of the wells during processing. If the hybridization is likely to exceed one hour, we also recommend the use of a dedicated hyb chamber. A suitable product is available on request from SCHOTT.
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Hyb chamber from Schott
(available on request)
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Can the MTP kit be used more than once?
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The MTP-96 coated plate, and superstructures are designed for single use only, and should not be re-used. However, the black plastic plate holder and pins may be cleaned and reused with a new coated plate and superstructure. If not all 96-wells are required for an assay then we recommend using the 16-well Nexterion® MPX-16 slides instead. These multi-well slides are available with the same coatings as the MTP-96 plates.
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Contact
SCHOTT Technical Glass
Solutions GmbH
Otto-Schott-Strasse 13 07745 Jena Germany
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