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MagNA Lyser Instrument Performance Data

 

Read in this article:

Processing Conditions
Eliminate Sensitivity Barriers with Increased Sample Input


Processing Conditions

Refer to the following tables for guidelines on setting up your homogenization:

Sample Material
(10 mg)*
Time Settings
(seconds)
Cooling
(between the runs in seconds)
Speed Average Yield
(µg)***
Average Purity
(OD 280/260 nm)***
Spleen 2 x 25 90 6,000 30 - 40 1.9
Liver 25 - 6,000 16 - 18 1.8
Lung 2 x 25 90 6,000 25 1.8
Kidney 25 - 6,000 20 1.8
           
Maize leaves** 20 - 5,000 10 n.d.
Maize polenta** 20 - 5,000 10 n.d.
Tortilla chips** 20 - 5,000 1 n.d.

Table 1: DNA
* Aliquot containing 10 mg sample material (here mouse and food samples) was taken for the DNA purification using the MagNA Pure LC DNA Isolation Kit II (Tissue).
** Centrifugation after the homogenization for 5 minutes at 2,200 x g.
*** Yield and purity strongly depend on the condition of the sample material.
n.d. not determined

Data kindly provided by Dr. Peterhänsel, RWTH Aachen, Germany.

 

 

Figure 1: Gel electrophoresis from genomic DNA isolated from tissue homogenized with the MagNA Lyser Instrument, using the MagNA Pure LC DNA Kit II (Tissue). Marker: DNA Marker III.

 

Sample Material
(10 mg)*
Time Settings
(cycles/ seconds)
Cooling
(between/after the runs in seconds)
Speed Average Yield
(µg total RNA)**
Average Purity
(OD 280/260 nm)**
Spleen 2 x 25 90 6,500 - 7,000 30 - 40 1.9
Liver 50 - 6,500 - 7,000 13 - 17 2.0
Thymoid tissue 60 90 6,500 n.d. n.d.
Heart 60 90 6,500 n.d. n.d.
Abdominal fat 60 90 6,500 n.d. n.d.
Aorta 60 90 6,500 n.d. n.d.
Other samples 1+n x 50 90 6,500 - 7,000 - -

Table 2: RNA/mRNA

* Aliquot containing 10 mg sample material (here mouse and human research samples) was taken to purify RNA either with the MagNA Pure LC RNA Isolation Kit III (Tissue) or the MagNA Pure LC mRNA Isolation Kit II (Tissue) homogenized with the MagNA Lyser Instrument.
** Yield and purity strongly depend on the condition of the sample material. The yield for mRNA was not determined.
n.d. not determined

Figure 2: Gel electrophoresis from total RNA isolated from tissue homogenized with the MagNA Lyser Instrument, using the MagNA Pure LC RNA Isolation Kit III (Tissue).

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Eliminate Sensitivity Barriers with Increased Sample Input

Rarely expressed targets in small numbers of target cells, as seen in experiments about minimal residual diseases, are difficult to detect. Increasing the cell number can improve sensitivity and lead to accurate results. Without the MagNA Lyser Instrument preprocessing, the MagNA Pure LC mRNA HS Kit can efficiently obtain mRNA from a maximum of 1 x 107 white blood cells (WBCs), as shown in research studies with human samples. However, using greater cell numbers results in a saturation effect with quantitative assays (see Figure 3). Homogenization of the lysate with the MagNA Lyser Instrument prior to the purification eliminates the amplification saturation at 1 x 107 cells and allows the use of up to 2.5 x 107 WBCs (see Figures 4 and 5), enhancing the analytical sensitivity of the assay.

   
Figure 3: mRNA was purified from different amounts of human white blood cells with the MagNA Pure LC mRNA HS Kit. G6PDH was amplified using the LightCycler® t(9;22) Quantification Kit.

 

Figure 4: mRNA was purified from different amounts of human white blood cells with the MagNA Pure LC mRNA HS Kit. The lysates from 2.5 x 107 cells and 5 x 107 cells were homogenized with the MagNA Lyser Instrument (2 x 50 seconds with 90 seconds cooling in between) prior to the mRNA purification. G6PDH was amplified using the LightCycler® t(9;22) Quantification Kit.

 

Figure 5: Scalability from 1 x 106 cells to 2.5 x 107 cells is represented in the graph and the table of the relationship between crossing points and cell numbers. The limitation of cell input is indicated by no change in crossing point with increased cell number.

 

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