Since we release TL2010s automated tissue mill, we continued to improve and explore the usage on different material. Recently, we conducted a test at a diagnostic reagent manufacturer company in Beijing， comparing the effectiveness and efficiency against traditional manual pestle.
Experiment objective
Compare manual grind and tissue mill grind in manner of efficiency and effectiveness. Evaluate feasibility of using TL2010s for extracting RNA from given sample.
Sample and reagent
1. Sample: 10 portions of fresh tissue drained to dry with tissue paper. Weigh and averagely divided into 2 tubes per portion, which is 20mg±0.3mg per tube.
Operation
1. Grind
[TL2010s automated grind]
Insert tube in adapter rack, add 300μl lysis buffer to sample, freeze sample in liquid nitrogen.
Add 2 ?3mm beads into frozen tube. Load adapter racks upon TL2010s. Set speed and time to 5 cycles, 60s/cycle, 10s pause between every cycle. After grinding, add 300μl more lysis buffer, make whole lysis buffer 600μl per tube.
[Manual] add sample and liquid nitrogen in mortar, grind sample with pestle immediately. Transfer sample powder in tubes add 600μl lysis buffer after weighing.
2. Preserve
Nucleic acid extraction, examine and preserve.
Extract DNA/RNA with kit; define purity and density by micro spectrophotometer. Preserve DNA at -20℃, RNA at -80℃.
Result
|
Sample |
DNA extraction |
RNA extraction |
||||
|
Sample No |
Grind method |
Sample loss |
Density(ug/ul) |
Purity A260/A280 |
Density ug/ul) |
Purity A260/A280 |
|
1 |
Automated |
0% |
349.8 |
2.07 |
157.4 |
1.91 |
|
Manual |
38% |
296.4 |
2.07 |
36.3 |
1.93 |
|
|
2 |
Automated |
0% |
1050.2 |
2.05 |
243.3 |
1.87 |
|
Manual |
44% |
188.2 |
2.10 |
199.4 |
1.87 |
|
|
3 |
Automated |
0% |
814.0 |
2.05 |
180.8 |
1.85 |
|
Manual |
40% |
137.8 |
2.08 |
210.9 |
1.86 |
|
|
4 |
Automated |
0% |
336.5 |
2.02 |
279.7 |
1.86 |
|
Manual |
11% |
236.5 |
2.07 |
210.2 |
1.87 |
|
|
5 |
Automated |
0% |
59.5 |
2.10 |
67.7 |
1.85 |
|
Manual |
44% |
44.4 |
2.09 |
11.0 |
1.80 |
|
|
6 |
Automated |
0% |
7.1 |
1.18 |
2.2 |
1.81 |
|
Manual |
60% |
7.1 |
1.56 |
6.0 |
1.11 |
|
|
7 |
Automated |
0% |
610.4 |
1.97 |
307.9 |
1.85 |
|
Manual |
18% |
475.8 |
2.01 |
507.5 |
1.86 |
|
|
8 |
Automated |
0% |
9.5 |
1.26 |
4.8 |
2.48 |
|
Manual |
61% |
34.2 |
1.32 |
2.9 |
1.82 |
|
|
9 |
Automated |
0% |
1.9 |
2.25 |
3.3 |
4.13 |
|
Manual |
44% |
7.0 |
1.89 |
1.6 |
2.68 |
|
|
10 |
Automated |
0% |
4.3 |
1.31 |
2.7 |
1.78 |
|
Manual |
48% |
6.2 |
1.26 |
11.8 |
1.93 |
|
Conclusion
There are no significant differences in purity and density of DNA, RNA extracted from fresh tissue, which means nucleic acid is not harmed during oscillation. However, there is significant sample loss of manual grind, comparing to 0 sample loss of automated grind, the advantage of Tissue mill 2010s is quite remarkable. Moreover in manner of time efficiency and contamination, manual grind requires complicate operation, result in risk of contamination and time waste.
|