The Reference Pipette is resistant against 6% nitric acid. The Tip ejector as well as the seals and o-rings might be affected by longer directly contact (spare parts available).

No. But the entire pipette can be autoclaved at 121 °C.

Research pipette Increment (µl) 0.1 - 2.5 0.002 0.5 - 10 0.01 2 - 20 0.02 10 - 100 0.1 20 - 200 0.2 30 - 300 0.2 100 - 1000 1 500 - 5000 5 1000 - 10000 10

Yes, the material of the pipette is resistant to toluene.

Yes, after cleaning, the piston should be re-greased. A special grease is supplied to enable you to do this.

We recommend to use a Reference RNA which should be chosen to have genes expressed in a reasonable way which you would like to study (to get quantitative data and not have a saturated signal or one which is too low), e.g. Stratagene Universal reference RNA (human # 740000; mouse # 740100; rat # 740200). A Reference RNA is a tool to control better experimental variance after RNA preparation.

• Compute ratios of your experimental samples with the reference RNA (do not use the housekeeping genes for normalization in the 2nd step as they are likely to be very different between experimental sample and reference sample).
• Then decide if the status (not in linear range, qualitative, quantitative / not the significance values of the ratios (unchanged, significant, highly significant) has to be incorporated somehow in the data analysis (like omitting the data which is not quantitative).
• Clustering techniques may then be applied and statistical tests on groups like ANOVA.

The proof of the signals from the hybridized, radioactive or alternative labeled molecules (->sample / target). By localization of the signals on the microarray the researcher gets the sequence information about the target material. A signal shows that the sequence of the target is complementary to the coupled and immobilized probe sequence spotted on the microarray. The detection method depends upon the type of labeling.

Yes.

200-400 bases.

The process of pairing together complementary strands of nucleic acids. The two strands are called hybrids.

60 °C if using the recommended hybridization buffer. If using other hybridization buffers the optimal temperature might change.

When using the Eppendorf hybridization chambers of the DualChip, the recommended volume is 100 µl.

For the RNA isolation, using a combination of a phenol-containing step (like Trizol® (Invitrogen) or QIAzol (Qiagen)) and a clean-up-column (like Qiagen RNeasy MinElute kit) is recommended.

Each gene on a DualChip microarray is represented by proprietary Xmer probes. These long DNA molecules combine the excellent hybridization efficiency of a cDNA fragment with the ability of an oligonucleotide to distinguish homologous genes. Xmers are perfect for quantitative and sensitive expression profiling experiments where it becomes necessary to study highly homologous genes.

It's unlikely that the pipette seal is faulty. More likely is that the tip seal is faulty. If the pipette is pressed down too hard when attaching tips from the rack, the leakage seal of the piston in the tip can be negatively affected. This is considerably more noticeable with highly viscous solutions than with fluid solutions. In addition, the tips may not be autoclaved.

No, the normalization process is based on the Internal Standards which are spotted on the DualChip as well as on the selected housekeeping genes.

BioAnalyzer (Agilent): RIN factor > 7
OD260/280 in TE-buffer at least 1.9
denaturating agarose gel (The overall quality of an RNA preparation may be assessed by electrophoresis on a denaturing 1% agarose gel (SamBrook et al., eds. (1989) Molecular cloning – a laboratory manual, 2nd ed. Cold spring Harbor, NY: Cold spring Harbor Laboratory Press). The integrity of the total RNA is assessed by visualization of intact ribosomal RNA bands. For the total RNA from higher eucaryotes, the size of the ribosomal bands should be 1.9 kb for the 18S-RNA and 4.7 kb for the 28S-RNA.) Concentration 0,35 – 1,6 µg/ µl (reaction volume: 6 µl for RNA)

The accession numbers for each gene on the DualChip include the version number of sequence. This version number defines the sequence which was used to generate the Xmer probe. NM_004324.1 is the ref-seq-code for the gene BAX, based on the first version. For ABI2, the ref-seq-code NM_005759.3 was used, i.e. the third version of the sequence was used to generate the Xmer sequence.
For specific sequences of Xmer-probes, please contact the microarray application support.

When slides are placed so that you can read “DualChip microarrays” and the barcode number, the spotted side will be up.

No.

The DualChip is available for human, mouse, and rat.

No, for the DualChips for gene expression, the Thermomixer comfort with the Thermoblock for slides DC is recommended to receive optimal results. There is no need to use expensive hybridization systems. Additionally, the Thermomixer can be used for the RT by using the Thermoblock for 1.5 ml tubes.

We clearly recommend to use the supplied washing buffer.

Surface of a solid slide (e.g. glass, silicon, plastic), on which systematically arranged biomolecular probes like nucleic acids, antibodies, peptides or proteins are immobilized.

Biomolecule of known identity (e.g. nucleic acid) which is immobilized on the slide. (Cf. conventional blotting methods, where probe = marked molecule, not immobilized). Nomenclature of probes and targets: The first publication dealing with microarray analysis (Schena et al., 1995) uses the term target to describe the immobilized DNA molecule onto the microarray substrate and probe to describe the sample that will be analyzed. This naming is based on the traditional northern blot nomenclature, where the term probe describes the immobilized sample, that will be analyzed. In principle the intention of northern blot analysis are the same as of microarray experiments, that is to measure the expression level of genes. However, the inversion of the physical locations of probes and targets led to some confusion within the microarray community. Therefore, you will always find some researchers, that are using probe to describe the immobilized DNA, whereas others describing with this term the sample, that will be analyzed. To stop this never-ending discussion, the editors of a comprehensive review of microarray technology entitled "The Chipping Forecast", published in Nature Genetics (1999), encouraged their authors to describe the immobilized DNA molecules always as probe and the free nucleic acid as target. Corresponding to this standard nomenclature, you will find this terminology within all Eppendorf writings concerning microarray technology.

Surface-activated carrier (e.g. slide made of glass or plastic) on which biomolecules (in this case nucleic acids) can be immobilized.

The kit should be stored at +4°C. The Internal Standard (2 tubes) should be stored at -80°C. After resuspension, the positive hybridization control should be stored at -20°C.

The Eppendorf microarray slides have the standard slide format of 25 x 75,4 mm. After hybridization, the hybridization frames are removed and the slide can be scanned.
The type of the scanner depends on the labeling strategy:
When using fluorescence labeling, every microarray slide reader is usable like scanners from Tecan, Molecular Devices/ Axon, or Perkin Elmer.
The output format should be a 16-bit *.tif-file.

"The pistons in the multi-channel bottom parts of the Research pro are connected to the top part by means of magnetic contact. This technique involves considerable benefits for maintaining and autoclaving the bottom part of the Research pro. If the pistons in the bottom part are torn away from the magnet, they no longer work and consequently no liquid can be aspirated by the tip. The cause of this might be sharp, jerky and frequent ""re-attaching"" when attaching the tips. Or the Research pro has been accidentally dropped. We would be pleased to let you know the measures you can take to remedy this problem - just contact our Application Hotline: support@eppendorf.com "

Yes, microtiter plates can be filled using the 8-channel Research pipette. The spacing between nose cones is 9.1 mm, corresponding to the dimension between holes from well to well of MTP and PCR plates.

Yes. Because every channel functions individually, the coincidental measurement deviation (imprecision) of the pipette is not influenced.

Yes. When setting the dispensing volume, the maximum number of dispensing procedures is shown on the right side of the display. In this case for 25 µl this would be 12 times and 10 times for 30 µl.

The O-rings of the pipette cones should be exchanged regularly. This is the only way to attach and eject the tips easily.

PICASO 2.2xxx can be operated in German, English or French. Spanish, Portuguese or Italian can additionally be installed for printing out documentation.

No. PICASO could be installed on several PCs of a network . The data bases are deposited on the server. The different positions have access to one common database although only one at a time. The access to the data files is selected via PICASO.

The operating instructions are hidden behind the CD cover.

If too many programs are open when the PC is started up (in one instance, there were 15), it is possible for them to interfere with one another and cause errors. Please remove some programs from Autostart and reinstall PICASO.

Yes, this is possible, i.e. the stored data (protocols) of the old version are definitely retained.

No, only one balance can be connected.

The key default settings for the balance are:

• baud rate: 2,400 baud
• data bits: 7
• stop bits: 2
• parity: even

A new pipette can be included in the inventory list under "in use at". To do this, enter the name of the institution where the pipette will be used. When you call up the inventory list you will see the "in use at" field in the title bar. By clicking on the field you will open a window in which you can choose the previously entered institution name under "Selection". Please confirm with "Yes". You will then be provided with an inventory list containing only those pipettes for the selected location / lab.

There are not yet any testing instructions available for this pipette. Testing instructions containing the name of the manufacturer, the volume and the tolerances must be created. To do this, call up the "Measuring / Test plans" menu. Select the menue "Edit / New" to create a new test plan. A pipette must be generated after creating and saving the new test plan. To do this, click on the new test plan (the last in the list) and select the menue "Edit / new pipette". Enter and save the data for the new pipette.

1. Select "Configuration / System / Password" in the menu bar. Activate password by entering PICASO as the password. Click on the floppy disk symbol to save the entry. Return to the menu bar.
2. Unlock tables by selecting "File / Maintenance / Tables" unlock.
This function removes network blocks which have inadvertently been set in the tables.

The EN/ISO 8655 recommend 10 measurements per volume for the calibration of piston stroke pipettes. This number of dispensings, like the evaluation limits, can also be changed.

Yes. The same formulae are used for calculation in all versions.

The Z-factor is automatically calculated by the program when the temperature and air pressure (of the solution to be measured) is entered.

No balance has been selected. Open the "Configuration / Balance / Manufacturer and type" menu and select a balance; save the entry by clicking on the floppy disk icon.

It uses "n-1".

This is possible only directly during measuring, not subsequently. It is possible to delete only the last measured value in each case, with both manual input and with automatic inclusion of values. This value then appears in brackets in the measuring protocol and is not included in the evaluation.

Yes. To open the protocol list either click on the protocol icon on the PICASO menu bar or use "Measuring / Reports". Now select the desired pipette. Clicking on the printer icon on the toolbar or use "Edit / Print" causes the selected data to be printed.

When installing, it is essential to ensure that the installation takes place in english, as otherwise the measuring protocol appears in german. Please reinstall the database.

The moisture trap is required for weighings of volumes <50 µl. Its use is independent of whether the evaluation is made with PICASO or not. The order number is 3113 809.000.

The volume setting was set to position 1 for both Multipettes. 48 dispensing steps (measuring values) were given for the Multipette and 50 for the Multipette plus. Testing can be carried out with any Combitip size. Information on testing and the limits of the ISO 8655 are also found in the SOP for Liquid Handling.

Both pipettes are tested according to the same DIN or ISO standard. However since handling of the two dispensers can vary, the test conditions are also different. For example the Multipette pro can be completely emptied in one step whereas the M. plus cannot (this is in pipette mode of the MP pro). Therefore the M. pro is tested with the total volume of the fitted Combitip and 1/10 of this volume. Dispensers that cannot be emptied in one step are tested with other volumes.

No, there is no multichannel adapter for the Multipette plus or pro.

No, there does not exist an 8-channel version for the Multipette plus, only for the old model 4780. Pipettes with 8 or 12 channels are only available for the Research pro or the Research multichannel.

Yes.

These chemicals damage the Multipette plus through contamination ( = incorrect use) and if immediate measures are not undertaken! With long-term use, damage may occur from the vapors. Short-term use (i.e. use for a couple of hours per day) will probably not cause damage to the device: the probability that no damage will occur is "very high" but unknown.

Yes, this is possible. However, it is important to be aware that the agar may set in the Combitip/plus if the solution is not dispensed quickly enough. Of course, these applications lie outside the scope of applications defined by Eppendorf, with the result that the specifications given cannot be guaranteed.

Since we do not know what the composition of this solution is we are unfortunately unable to answer this question. To disinfect a Multipette we recommend the use of 60 % isopropanol. Please take care that no solution enters the Multipette or comes onto the display.

The Multipette plus can only be tested by electronic methods. In the event of any problems with the multipette, it should be sent to the appropriate Eppendorf service partner or the Eppendorf Marketing office for testing.

No, only our battery (4980.215.003) can be used. One reason for this is excessive current protection, for example.

No, the Multipette can also be used with a Combitip that has not been completely filled. In this case please bear in mind that the first dispensing step has to be discarded!

The batch number is engraved directly under the dispensing lever or in the battery chamber.

"Yes, pore widths of 0.3 µm and 0.2 µm may also be used. This has only a minimal effect on aspirating and dispensing speeds. When purchasing, please note the shape of the filter connections; double-sided Male Luer Slip (MLS)."

Please make certain that the pipette is pushed far enough into the adapter of Easypet. It could be the case that the adapter or the pipette is damaged.

Reduced suction capacity may be caused by a wetted membrane filter or a discharged battery.

Yes. The device is even recharged during the pipetting process.

Nickel-cadmium batteries are used in Easypet.

A fully charged battery can be used for approximately 7 hours.

No. However, there are also no batteries with the dimensions of the 3 stipulated batteries.

The 0.45 µm filter installed in Easypet is not resistant to the named reagents. Please replace the filter with a new 0.2 µm filter, which is resistant to the named solutions.

Liquids which give off vapors that attack silicon, NBR, ABS (Acryl Nitrile-Butadene-Styrol-Copolymer), POM (Polyoxymethylene) or PVDF (Polyvinylidenfluoride).

This depends on the dispensing technique used. For liquid dispensing without a pump, it is possible to carry out 2,000 cycles. With a pump, only 650 are possible.

The wall holder has a Velcro strip that is pressed onto the wall after the protective film has been removed. The wall holder can be pulled off sideways from the Velcro.

For both charging stand the same power supply unit with 7,5 V and 800 mA is to be used. The order no. is: 0013 613.370.

The order number for the nickel-metal hydride rechargeable batteries is 4860 501.002

Yes, this is possible. For example, it is important if you are intending working with Filtertips. The volume limitation you can find in the operating manual, sec. 5.11.

The RNS option = Rinse means pipetting with subsequent rinse feed. Rinse feed is recommended for volumes < 10 µl. and is suitable for larger volumes when mixing the dispensing volume and the prescribed liquid. The setting "max. speed" is often recommendable. Feed rinsing or mixing is always carried out with the previously set dispensing volume.

Compared to the multi-channel pipette, the single-channel Research pro 20 - 300 µl pipette has slight differences at the slowest (for gel loading) and fastest speeds. However, this has no effect on the "Mix" function.

Aspirating 5 ml water at the fastest speed takes 1.25 sec.

No, the Research pro does not fit in the pipette carousel, only in the charging stand designed for it.

The Research pro pipette is stable to UV. In the event of prolonged irradiation with UV light, material discolorations may occur, but these do not affect the proper function of the pipette.

As dibutylamine is very similar to ammonium, the Varispenser is suitable here.

Eppendorf UserGuide no. 17 contains a list of chemical stability. We are happy to send you this list upon request.

"Yes. Due to the hygroscopic properties of morpholine, please use the drying tube (4960 851.00). The drying tube should be filled with dried calcium carbonate. "

The silicone of the sealing ring may be attacked after prolonged contact with acetonitrile. However, this has no effect on the precision of dispensing.

Yes, you can use the Varispenser/plus for this application.

Dibutylamine is very similar to ammonia. The Varispenser is resistant to dibutylamine.

Yes, the Varispensers are not chemically attacked by iodine solution. However, iodine solution in high concentrations may precipitate and block valves.

Although neither 0,1 N butylamine nor ethyl acetate were tested explicitly, it should be possible to use them.

"Varispensers are measurement devices for which the guaranteed accuracy, precision or error limits only apply in a restricted temperature range of 15 - 40 °C (conforming to DIN 12650). Applications with liquids outside this temperature range can only be carried out at the user's own risk. At higher temperatures up to 60°C the plastic materials PTFE and polypropylene will be fully chemically resistant, but mechanically they will become slightly softer. Therefore the Varispenser may only be handled with a moderate use of force. When hot agar is aspirated out of the bottle, cooling will take place in the cold Varispenser. Since agar becomes much firmer at ca. 35 °C, blockage can easily occur. In any case immediate cleaning will be required, in particular of the valve. As an alternative the Varipenser as a whole can be warmed at 60 °C before it is used. We have no data for dispensing agar at 55 °C."

We cannot recommend this application as we have not tested it thoroughly. It is possible that vapors or splashes of the solution might damage the Varispenser on heating. It is furthermore impossible to ensure that the Varispenser is successfully sterilized, as even if it is only loosely on the flask, sufficient hot steam can probably not penetrate it.

The Varispenser should be checked for leakproofness and function after approximately 20 autoclavings. Note: The Varispenser may not be taken apart for autoclaving.

The sealing ring is made of silicone (covered with a PTFE washer).

Yes.

No, that is not possible. For this application an attachment would have to be screwed onto the discharge tube at the front, something which is not possible with our Varispensers.

An absolutely normal PP adapter can be used as Alsever's solution is harmless; it is used as anticoagulant for the storage of test erythrocytes. Content of solution: 2.05% glucose, 0.42% NaCl, 0.8% trisodium citrate, 0.055% citric acid in dist. water.

For the Varispenser/Varispenser plus 0.5-2.5 ml //1-5 ml // 2-10 ml the adapter order no. 4960 800.066 (thread 32->38 mm) will fit, and for the Varispenser/Varispenser plus 5-25 ml // 10-50 ml // 20-100 ml the adapter order no. 4960 800.155 (thread 45->38 mm).

Yes, this is possible. The Top Buret has both the chemical resistance and the relevant dispensing characteristics for this task.

A dispenser, such as the Eppendorf Varispenser, delivers a preset volume. A reciprocating pump is used to draw up the required volume and deliver it at once. This process is particularly suitable for identical volumes which are dispensed frequently. With a TopBuret an unknown volume is delivered. With analytical procedures such as the determination of a pH value or titration, liquid is supplied until the required effect occurs. The quantity of liquid delivered is then read off at the burette. During dispensing the liquid is not delivered from a reservoir but is supplied continuously from the bottle.

Yes. If the voltage is too low (or if the battery is flat), a battery symbol lights up in top left of the LCD. The battery should then be changed.

The difference is the amount of liquid that can be dispensed by turning the dispensing wheel one revolution. It is possible to dispense 2.5 ml per revolution with Top Buret M, and 5 ml per revolution with Top Buret H.

The maximum volume pro time unit that can be dispensed (limited by the flow resistance) is 1.25 ml/sec for the TopBuret M and 2.5 ml/sec for the TopBuret H, since one revolution should not be faster than 2 sec for reasons of accuracy. 2.5 ml/revolution can be dispensed with the TopBuret M and 5 ml/revolution for the TopBuret H.

Yes.

The device as a whole cannot be autoclaved, but the telescopic filling tube made of PTFE can be autoclaved up to the permitted temperature of 121 °C. It is important not to separate the two parts of the tube for autoclaving and to wait for the tube to cool down completely before inserting it back in the Top Buret.

Yes. This certificate is on the back page of the operating manual.

Eppendorf Application no. 6 (3/95) contains a list of chemical stability. We are happy to send you this list upon request.

Yes

Although neither 0,1 N butylamine nor ethyl acetate were tested explicitly, it should be possible to use them.

Yes. Please use our PTFE adapter to do so.

No. To disinfect the Multipettes, we recommend wiping the device with a cloth soaked in disinfectant (e.g. isopropanol). When doing so, please ensure that no liquid enters the device or the display. Sterility can only be guaranteed by the use of sterile Biopur Combitips plus.

The Multipette stream / Xstream works with an even reproducible rhythm during pipetting. This has a positive effect on the random error (imprecision).

The Multipette stream features the functions dispensing, automatic dispensing and pipetting. The Multipette Xstream includes additional features sequential dispensing, aspiration and titration. Both repeater use identical Combitips plus.

Main features are: very low weight, spring loaded tip cone (exception: 5 mL and 10 mL Research plus) which leads to an easy and homogeneous tip attachment even when working with a multi-channel pipette and thus, to very low operation forces and finally, an easy adjustment by the user. In addition, the Research plus shows a very good accuracy and precision, is fully autoclavable at 121 °C, 1 bar for 20 min, features excellent chemical resistance and has an adjustment display offering optical control if an adjustment of the pipette has been performed.

Yes. Please notice that the usage of aggressive chemicals over a longer period of time will reduce the life span of the pipette.

It does not affect the accuracy of pipetting if the Research plus multi-channel pipette is not fully attached with tips or if individual channels are removed.

Yes. Please notice that the usage of aggressive chemicals like hydrochloric acid over a longer period of time will reduce the life span of the pipette.

No, it is not necessary.

Yes.

If the pipette is contaminated with a liquid which enters the pipette's internal parts, the user will have to disassemble the lower part, clean the piston and the nose cone . In this case, the grease is removed from the piston and the piston needs to be regreased before reassembly. To do so, we recommend to use our pipette grease (1 g, order no. 0013 063.010).

The user manual of the Research plus mentions specific settings for some volumes of the most common liquid classes. These values are for orientation only as the systematic and random error are affected by the mode of operation, the tip used and other factors like the temperature. It is essential to check these data according to your own work method.

Vapor will penetrate the inside of the pipette during autoclaving but this will not damage anything. We tested our pipettes with 175 autoclaving cycles (121 °C, 1 bar, 20 min). The number of respective tests chosen corresponds to an average of 5 years of use.

For technical reasons, the results are generally more precise when a volume setting is carried out from a higher down to a lower value. This is valid for all manual pipettes.

We recommend to wet a cloth with water or 70% isopropanol and to remove the contamination.

As each Research plus pipette has been calibrated and adjusted before delivery, the adjustment display shows "0". As soon as the adjustment is changed by the user, the reading on the display will be changed, too. Therefore, the adjustment display provides optical control if a pipette has been adjusted to another volume.

For dispensing tasks in which a pipette tip is used over a long time or is slightly bended, it can be helpful to deactivate the spring loading of the tip cone by installing the locking ring. This can also be helpful for pipette tips which are not designed for the Eppendorf pipette cones.

"Yes, the Xplorer is resistant to solvents. Only the display cannot come into contact with the solvent."

"Yes, there are several options for adjusting the pipette in the Options operating mode, e.g. to ethanol 75 %25 or gylcerol 50 %25. Furthermore, individual adjustments are possible, as 1-point, 2-point or 3-point settings."

Yes, the volume of the acustic feedback can be set to the levels 0-8 in the Options operating mode.

This is independent from the aspiration volume. Please contact the Application Hotline at www.support@eppendorf.com if you have any further questions.

"The Xplorer is UV-stable. Longer periods of irradiation can lead to very minimal material discoloration that does not have any effect on functionality."

Yes, a volume limit can be defined in the "Options" operating mode.

"No, the Xplorer cannot be charged in the Multipette charging stand. But the charging shells of the Xplorer charging stand can be exchanged. A charging shell for a Multipette stream/Xstream can be used instead of a charging shell for an Xplorer. "

Yes the battery can be individually ordered by customers as a spare part, or reordered.

In the operating mode "Pipetting and mixing P/M," the sample is automatically mixed with the preparation after dispensing. The mixing cycles and mixing volume can be individually defined for this.

On the Xplorer, the pipette can be adjusted to different liquids or geographic heights using 1, 2 or 3-point adjustment, taking into account the corresponding density and associated scale results. The setting for ethanol 75 %25 or glycerol 50 %25 is completed using an internally determined factor that takes into account the density.