Top

FrameStar® PCR plates

This product is covered by one or more of the following U.S. patents or their foreign counterparts, owned by Eppendorf AG: U.S. Patent Nos. 7,347,977 and 6,340,589.

Our FrameStar® two-component technology reduces evaporation from PCR plates, improving your results and allowing you to reduce the volume of expensive reagents to save you money. FrameStar® plates combine the advantages of thin-walled polypropylene tubes for optimum PCR results, with a rigid polycarbonate frame to increase the thermal stability of the plate during the PCR process.

This means that compared to a standard polypropylene plate, the FrameStar® plate will not warp and expand during rapid heating and cooling, meaning the integrity of seals will remain consistent and therefore less of your sample will evaporate. This in turn means you can spend less money on expensive reagents, as you are retaining more during the PCR process. Read more ▼

FrameStar® PCR plates

This product is covered by one or more of the following U.S. patents or their foreign counterparts, owned by Eppendorf AG: U.S. Patent Nos. 7,347,977 and 6,340,589.

Our FrameStar® two-component technology reduces evaporation from PCR plates, improving your results and allowing you to reduce the volume of expensive reagents to save you money. FrameStar® plates combine the advantages of thin-walled polypropylene tubes for optimum PCR results, with a rigid polycarbonate frame to increase the thermal stability of the plate during the PCR process.

This means that compared to a standard polypropylene plate, the FrameStar® plate will not warp and expand during rapid heating and cooling, meaning the integrity of seals will remain consistent and therefore less of your sample will evaporate. This in turn means you can spend less money on expensive reagents, as you are retaining more during the PCR process.

FrameStar® PCR plates maximize thermal stability at high temperatures, which prevents sample loss by minimizing thermal expansion during PCR. The two-component design combines the advantages of thin wall polypropylene tubes for optimum PCR results, and a rigid polycarbonate skirt and deck for the highest thermal stability and rigidity. In contrast to standard one-piece PCR plates, evaporation from the corner positions of the plate and outer rows of wells is minimal, which allows for the downscaling of reagent volumes and therefore saves costs.

Why choose FrameStar® PCR plates

  • Two-component technology reduces thermal expansion and sample evaporation
  • Reduction of evaporation leads to improved consistency of PCR results
  • Well spacing and position post-PCR remain accurate so liquid handling devices can reliably add or remove the smallest quantities from the plate
  • Ideal for robotics, as plate distortion is eliminated post-PCR
  • FrameStar® is ideal for assay miniaturization due to rigid skirt eliminating warping, giving better sealing properties
  • Cost saving due to downscaling of reaction volumes
  • Lack of warpage reduces variation of fluorescent signals in optical assays, such as qPCR

Thermal expansion and sample evaporation

Figure 1: Standard plates with polypropylene frame expand by up to 2 mm during thermal cycling, which leads to movement of wells away from the plate centre. This movement is most significant in the corner positions and outer rows of the plate. Sealing sheets do not expand at this rate so that the movement of wells will weaken the seal and lead to evaporation especially in corner positions and outer rows
Figure 1

In standard 1-piece PCR plates

PP is the optimum material for PCR tubes. It provides the most efficient heat transfer, as well as an inert surface with low binding capabilities for nucleic acids, proteins and other molecules. However, the material is not thermally stable in a plate format, and expands and contracts during each PCR cycle (Figure 1).

Such thermal expansion will weaken the plate seal and leads to sample evaporation mainly from corner wells and outer rows.

Thermal cycler blocks do not prevent thermal expansion of PCR plates. PCR blocks do not support PCR plates from the sides, and the high temperatures from the thermal block and heated lid accelerate expansion of the plates (Figure 2).

Figure 2: Side-on view of a PCR plate in a thermal cycler. The sealed plate is sandwiched between the cycler block and the heated lid, but it is only partly fixed in position at the bottom of tubes, allowing the plate to expand horizontally
Figure 2

Evaporation from one piece PP plates is highest in the outer wells. Since thermal expansion and movement of wells in one piece PP plates is enhanced around the edges of the plates (see Figure 1), evaporation is the highest from the two outer rows of wells.

Figure 3 illustrates the levels of sample evaporation from different areas of PP plates. only the inner 32 wells of a one piece 96 well plate show low levels of evaporation, but sample loss is high from the two outer rows which contain more than 65% of the wells.

In FrameStar® 2-component PCR plates

FrameStar® 2-component technology reduces thermal expansion and sample evaporation. The polycarbonate frame of FrameStar® plates is significantly more heat resistant than standard PP plates, which reduces thermal expansion to a minimum. For this reason the seal integrity remains intact even at elevated temperatures during PCR.

Figure 3: Evaporation from the outer rows (red) is highest, medium level evaporation occurs in the second row (yellow) and sample loss from the inner 32 wells is lowest
Figure 3

To illustrate this advantage of our two-component technology we have compared evaporation from one piece PP plates and FrameStar® PCR plates. Each well of a non-skirted 96 well plate (single piece, PP) and a Framestar® non-skirted design (code 4ti-0710) was filled with 10 μl H2O. The plates were sealed with a qPCR adhesive (code 4ti-0560) and the weight of plates was measured before and after performing PCR (30 cycles x 15’’ 95°C; 15’’ 55°C).

Table 1 shows that the average volume loss from one piece PP plates was 2.3 μl per well which is equal to 23% of the total reaction volume. in contrast the volume loss from Framestar® plates was only 0.49 μl per well using adhesive sealing. For easier sample identification, we supply all microplate designs with a printed alphanumeric grid reference.

Plate type FrameStar® plate (4ti-0710) 1-component plate
Starting weight 26.678 g 17.807 g
Weight post PCR 26.631 g 17.586 g
Weight loss 0.047 g 0.221 g
Volume loss total/well 47 µl/0.49 µl 221 µl/2.3 µl

Table 1: Weight and volume loss from 96 well PCR plates. Results shown are averages from 5 plates of each plate type. One piece PP plates showed more than 4 times higher volume loss than FrameStar® plates

Consistency of PCR results

We have compared the degree of evaporation from different areas of one piece PP and FrameStar® PCR plates. First, the 64 outer wells (two outer rows) of both plate types were filled with 10 μl H2O, plates were then sealed with a qPCR adhesive (code 4ti-0560) and their total weight determined before and after PCR. The experiment was repeated with a set of plates of which the inner 32 wells (green area in Figure 3 below) were filled. Table 2 shows that evaporation from outer wells of standard PP plates was significant and 65% higher than from inner wells. In contrast, FrameStar® plates showed low levels of sample loss across the plate.

The results show that reaction volumes remain consistent across the 96 wells (or 384 wells, data not shown) in FrameStar® plates. In contrast, the reaction volumes in standard plates will differ increasingly between wells during PCR. Buffer concentrations in outer rows will increase dramatically and result in reduced enzyme activity. In extreme cases samples will fully evaporate.

Plate type FrameStar® plate (4ti-0710) 1-component plate
Outer 64 wells
Starting weight 26.230 g 17.299 g
Weight post PCR 26.193 g 17.118 g
Weight loss 0.037 g 0.181 g
Volume loss total/well 37 µl/0.57 µl 181 µl/2.8 µl
Inner 32 wells
Starting weight 25.841 g 17.132 g
Weight post PCR 25.824 g 17.078 g
Weight loss 0.017 g 0.054 g
Volume loss total/well 17 µl/0.53 µl 54 µl/1.69 µl

Table 2: Weight and volume loss from different sections of 96 well PCR plates. Results shown are averages from plates of each plate type. Volume loss from the outer wells of one piece PP plates was 5-times higher than from FrameStar® plates

Reduction of assay volumes and cost

Figure 4: Comparable Ct values and mean deviations were obtained with 15 μl and 25 μl SyBr Green assays in heat sealed FrameStar® plates
Figure 4

Due to the much improved seal integrity, reaction volumes can often be reduced when using FrameStar® plates. Such downscaling of experiments can be successfully implemented without any loss of assay sensitivity or consistency (Figure 4) and reagent savings can be considerable.

Plate distortion post-PCR

Distortion tests have been performed with different PCR profiles using a variety of sealing materials. The results shown were obtained using the following protocol with 5 μl reaction volumes, plates sealed with adhesive sheets (4titude® code 4ti-0500), in a Thermo Px2 Cycler: 1 min 94 ºC, 30 x (30 sec 94 ºC, 30 sec 55 ºC, 30 sec 72 ºC), 5 min 72 ºC, holding step at 4 ºC.

Averages from measurements of 10 plate samples are shown. X- and Y- axis measurements were taken in the middle sections of the plate skirt. Warpage was measured as a maximum deviation from a flat plane. The results in Table 3 again demonstrate the high level of thermal stability of our FrameStar® plates compared to a standard one piece PP plate.

Plate type FrameStar® plate 1-component plate
96 well
X-axis 0.02 mm 1.18 mm
Y-axis 0.02 mm 1.69 mm
Warpage 0.03 mm 1.80 mm
384 wells
X-axis 0.02 mm 1.22 mm
Y-axis 0.02 mm 1.82 mm
Warpage 0.03 mm 2.20 mm

Table 3: Plate distortion post-PCR

Improved sealing

In addition, FrameStar® technology provides a much improved level of seal integrity: During PCR, with heat seals or adhesive seals, the excessive thermal expansion/contraction of standard one-piece polypropylene plates can have a damaging effect on seal quality.

This is especially evident around the edge of the plate where these forces are at their maximum resulting in significant sample evaporation from the outer wells. in contrast, the higher thermal stability of FrameStar® plates results in a reliable seal in all areas of the plate during PCR and in cold storage down to -80 ºC.

Reduced risk of contamination

Raised rims around each tube aid the prevention of cross contamination between samples.

Plate compatibility

FrameStar® plates are compatible with most PCR cyclers, real-time detection systems and sequencers. The plates have been successfully tested for reliable use on most automation platforms. Contact us for further details. Read less ▲