|A platform for high-throughput molecular characterization of recombinant monoclonal antibodies|
Mark J. Bailey, Andrew D. Hooker, Carolyn S. Adams, Shuhong Zhang, David C. James
J. of Chromatography B, 826 (2005) 177-187, 2005 2005
|Optimization of a simple, automatable extraction method to recover sufficient DNA from low copy number DNA samples for generation of short tandem repeat profiles.|
Schiffner LA, Bajda EJ, Prinz M, Sebestyen J, Shaler R, Caragine TA.
Croat Med J. 2005 Aug;46(4):578-86. 2005
|I am using the Multiscreen system and my samples won't flow through the plate. What's wrong?||There are a number of reasons why samples might not flow through the MultiScreen system. Be sure that the vacuum line is clear and not clogged. Be certain that your samples do not have a high particulate load. If they do, you may wish to dilute your samples in filtered buffers. Also, if you are working with cells, make sure that you do not have more than 10^6 cells in each well. If you are over this amount, dilute the cell suspension ot try working with a more open membrane ( larger pore size). Be certain that you do not have the lid on the Multiscreen system. this should be removed prior to filtering in the vacuum manifold. Finally , be sure that if you are not using a whole plate that you have sealed the empty wells with plate sealing tape.
In addition, ensure that the MultiScreen plate is forming a secure seal with the top gasket of the MultiScreen manifold. This can be checked by trying to lift the plate off the manifold with the vacuum on. If the seal is not secure press the four corners of the plate with the vacuum on to seal the plate.
|The Tech Note "TN053" entitled "Dye Terminator Removal and Sequencing Reaction Clean-up Using Multiscreen 96- Well Filtration Plates" states to add the sequencing reaction to the center of each well (already containing the prepared G-50 resin column). Can I add less than the standard 20ul volumes of sample to each column?||Yes. You can add less. When adding volumes less than 10ul you should prewet the resin.|
|What type of scintillant do you recommend for use in MultiScreen?||Although we don't recommend a specific type there are some guidelines. If the reading is done in the MultiScreen plate it important to use a scintilation fluid specific for microtiter plates. The standard scintillation fluid for reading in vials in higher volume and is mixed differently so it shouldn't be used with a microtiter plate.
MicroScint O is for organic and so it would be used with Durapore membrane. In our R&D labs, we have used MicroScint 20 and 40 interchangeably. Supermix (by Wallac) is a microtiter scintillant and has also been used interchangeably with MicroScint 20 and 40.
|What is the well depth and maximum volume capacity of a MultiScreen plate?||The well depth of a 96 well MultiScreen plate is 1.245 cm. The well depth for a 384 well MultiScreen plate is 1.2 cm. The maximum working volume of a 96 well plate is 300 ul. The maximum working volume for a 384 well plate is 100 ul.|
|When I incubate MultiScreen plates, liquid leaks from the bottom of the plate. Is there a way to prevent leakage from the plate?||Normally, the Multiscreen filtration plates do not leak when placed on a dry, non absorbent surface during incubation. If you are experiencing leakage, however, be certain that you have asked yourself the following questions:
1. Do I have a high surfactant concentration in the sample?
It is important that you not exceed .1% surfactant in each well.
2. Do I have organic solvents in my solution?
Although the Multiscreen plates have a limited tolerance to organic solevents it is important that you have verified that the concentration and type of solvent used in your assay is compatible with both the membrane and plate plastic. Otherwise, a solvent resistant plate may be more appropriate.
3. Is the underdrain coming in contact with an absorbent surface?
If you lay the plate on lab "chucks" it will cause liquid to wick out from the plate.
4. Have I inspected the plate bottom for microdroplets?
Be sure to blot the plate onto absorbent material prior to all incubations.
|I can't get any flow through the MultiScreen NA (lysate clearing plate) when performing the plasmid prep protocol. What's wrong?||Clogging of the plate is usually due to inappropriate culture medium resulting in overgrowth or lysis. Use 2x LB, not terrific broth or another excessively rich medium.|
|I am using the Multiscreen system and the Multiple Punch assembly for liquid scintillation counting and I am not getting any counts. What might be causing this?||Low or inconsistent counts most commonly results from the lack of release of the "labeled" material from the membrane. This is especially an issue with tritium. Millipore has developed a procedure for the release of radioactive label from membrane by adding 0.5 ml of dilute (0.42%) sodium hypochlorite to the scintillation vial containing the "punched" membrane and agitating for thirty minutes prior to the addition of the scintillation cocktail. Please request Technical Brief TB038 for additional details on this procedure.|