How to extract pork cells and what method to extract the cells

First, cut a small piece of pork, put it in a petri dish, then scrape some crumbs on the meat with a glass rod, apply it to a glass slide with saline, cover it with a coverslip, and you can observe it with a microscope, but it is best to stain it with a dye so that you can see it more clearly. As for the extraction of pork cells, just put them in the container you prepared instead of putting them on a glass slide.

First, the large particulate matter is filtered out with filter paper, and then centrifuged at low speed (200g) for 5 minutes, and the cells will be at the bottom of the tube. Aspirate the supernatant and resuspend the cells with saline and centrifuge again.

Filtration. Use a special filter paper.

It is possible to filter out things that are larger than the cells.

Smaller volumes than cells can pass through filter paper.

When isolating and purifying proteins with GST tags in vitro, only the fusion protein with GST can be specifically bound to the column during the GST pulldown process, and impurities such as nucleic acids will flow away with the effluent without interfering with your experimental results.

With a cell lysis kit, it should be possible, the imported one is more expensive, you can call ** to ask Nanjing KGI domestic box can be used.

(1) Aqueous solution extraction method.

The aqueous solution of dilute salt and buffer system has good stability and high solubility to protein, and is the most commonly used solvent for protein extraction, usually 1-5 times the volume of raw materials, and uniform stirring is required during extraction to facilitate the dissolution of protein. The temperature of extraction depends on the nature of the active ingredient. On the one hand, the solubility of most proteins increases with increasing temperature, so a high temperature is conducive to dissolution and shortens the extraction time.

On the other hand, the increase in temperature denatures and inactivates proteins, so it is common to use low temperatures (below 5 degrees Celsius) to extract proteins and enzymes based on this consideration. In order to avoid degradation during protein extraction, proteolytic enzyme inhibitors (such as diisopropyl fluorophosphate, iodoacetic acid, etc.) can be added.

The selection of pH and salt concentration of the extract is discussed below.

1. pH value protein, enzyme is amphoteric electrolyte with isoelectric point, and the pH value of the extract should be selected in the pH range on both sides of the isoelectric point. When extracting with dilute acid or dilute alkali, it should be prevented from changing the dissociable group of the protein caused by excessive acid or alkaline, which will lead to irreversible changes in the conformation of the protein.

2. The salt concentration can promote the dissolution of proteins, which is called salt dissolution. At the same time, the dilute salt solution has the advantage of protecting the protein from denaturation due to the partial combination of salt ions and proteins, so a small amount of neutral salts such as NaCl is added to the extract, generally in moles. liter concentration is appropriate.

Phosphate and carbonate isotonic salt solutions are often used in buffers.

2) Organic solvent extraction method.

Some proteins and enzymes that bind firmly to lipids or have more non-polar side chains in the molecule are insoluble in water, dilute salt solution, dilute acid or dilute alkali, and can be used with organic solvents such as ethanol, acetone and butanol, which have a certain hydrophilicity, as well as strong lipophilicity, and are ideal for the extraction of lipoprotein. However, it must be operated at low temperatures. The butanol extraction method is particularly superior to the extraction of some proteins and enzymes that are tightly bound to lipids, firstly, because butanol is highly lipophilic, especially the ability to dissolve phospholipids; Second, butanol is both hydrophilic, and in the solubility range (degree is 10%, 40 degrees will not cause denaturation and inactivation of enzymes.

The cell compartment is isolated and not extracted, which is very different.

You can extract DNA, because DNA is a relatively stable genetic material, if it is not too long, because of the large number of cells in the organism, you can always find a relatively complete fragment of DNA, and then copy this fragment, which can be used for research, but if the organism suffers from chemical interference, the difficulty of extracting DNA will be greatly increased.

If you want to prevent the extraction of important DNA fragments from the weight of four organisms, you can process the organisms.

Hydrolyze the cell membrane with phospholipase before centrifugation with a centrifuge.

Extracting the pellet, using phospholipase, protease, and nuclease, hydrolyze the nucleus, and then evaporate the liquid and cauterize a mixture of nitric acid and hydrochloric acid with low and high concentrations.

After the cell dies, the intracellular DNA will begin to degrade, first broken into one by one, until finally degraded by microorganisms into inorganic salts and returned to nature.

It can be dry, but not for too long.

Because after the leaves and branches dry up, the first thing to disintegrate is the DNA sequence and protein, so if it has been dry for a while, the extraction may only be DNA fragments, which is why modern humans cannot clone dinosaurs.

But it is still possible in a special state, such as freezing.

It can be extracted, but the DNA sequence in some dead cells is destroyed, and the extracted sequence may not be complete.

Dead branches and leaves are not all dead cells, and there are often some living cells left.

As long as the storage of the substance you are talking about is not DNA decomposition, it can be extracted.

It is possible to extract DNA, for example, the field of paleontology research is specialized in the study of DNA extracted from fossils. Specifically, it also depends on the degree to which the DNA in the cells of dead branches and leaves is degraded, and the DNA in the dead cells will definitely be degraded, but if the degradation is not severe (large fragment), then the target fragment can still be obtained by extraction and amplification; And if the degradation is very strong, it will be more difficult.

The difference with live cell extraction is that total DNA is extracted from live cells; Dead cells, on the other hand, may only have extracted a portion (fragment) of total DNA.

It's not absolute, it's generally very seriously degraded, and if you can, the technology is super powerful.

Cells are dead, but DNA never changes.

I suggest you take a look at the biology book for your sophomore year of high school!

Pork skin is rich in collagen, and it is possible to refine it, but the absorption effect is not very good.