Through a large number of sample crushing tests, Dingyuanyuan Technology found that animal tissue samples are more difficult to break than other samples, especially some visceral tissues, connective tissues, heart muscle tissues, and lung tissues. It is generally difficult to use manual or conventional tissue homogenizers. Completely broken. Because these animal tissue samples generally have high toughness, if manual grinding or tissue homogenizer is used, not only the amount of treatment is too small, it is easy to be contaminated and difficult to clean, and the sample after treatment is not uniform enough and detailed, and the subsequent extraction of RNA is not achieved. Requirements. From March to April 2015, a biological company in Beijing used TL2020 tissue grinding and crushing instrument (also known as TL2020 high-throughput tissue grinder) to grind visceral and tumor cells, and subsequently extracted RNA. The experiment was smooth and the effect was very good. The following is The main process of this grinding experiment can be used by relevant experimental personnel for reference:
ã€Material preparation】
· Sample types: human tumor cells, pancreas, prostate, fat, muscle, each tissue was ground 2 samples, and stored with liquid nitrogen.
· Types of grinding accessories: Ding Yiyuan TL2020 tissue grinding and crushing machine 2.0ml grinding tube adapter (28 × 2.0ml) 1 pair, 6mm stainless steel grinding beads.
[grinding experiment flow]
1. Pretreatment of 6mm stainless steel grinding beads by high temperature sterilization;
2. The sterilized grinding beads and the grinding tube adapter are placed in liquid nitrogen while pre-cooling;
3. Remove the sample from the liquid nitrogen and cut into small pieces of sample approximately 4-5 mm wide in a mortar.
4. Add 2 pieces of 6mm sterile round bottom centrifuge tube (import): 1 piece of pre-cooled 6mm grinding steel ball, 1 piece of cut sample block, and 1 piece of pre-cooled 6mm grinding steel ball.
5. After ensuring that there are 2 6mm ground steel balls and one sample block in the centrifuge tube, quickly cover the tube and place it symmetrically into the pre-cooled 2.0ml grinding tube adapter hole.
6. Fix the adapter on the two rocker arms of the TL2020 tissue grinding crusher, set the speed of 1800 rpm, time 120S (pancreas sample 180S), start grinding with one button.
8. At the end of the grinding time, unload the adapter to remove the centrifuge tube, quickly add the lysate to the tube, and follow the subsequent RNA extraction protocol.
[grinding effect]
• Subsequent successful extraction of RNA, and the electropherogram bands are clearly visible.
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Figure 1: TL2020 Tissue Breaker Real Machine Photo | Figure 2: Grinded sample in a centrifuge tube | Figure 3: Extracted RNA electropherogram |
In this experiment, TL2020 tissue grinding and crushing instrument was used to crush 56 pieces of 2ml visceral sample tissue at high speed at one time. It is not only flexible and convenient to operate, but also safe and easy to carry out liquid nitrogen freezing and grinding operation, ensuring temperature sensitivity of RNA extraction and subsequent experiments. Going smoothly. In addition, similar animal sample tissue grinding with TL2020 avoids cross-contamination between samples, eliminating the cumbersome work of cleaning instruments, and greatly improves the efficiency of experiments with large sample volumes and requiring aseptic processing.
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