Differences in whole blood, serum and plasma samples during DNA detection
Differences in whole blood, serum and plasma samples during DNA detection
Article source: Luoyang Jiente Biotechnology Co., Ltd.
Many blood DNA extraction kits can be applied to whole blood, plasma, and serum on applicable samples. Therefore, many teachers think that these three samples can be used when doing nucleic acid diagnostic experiments. However, the results obtained are often not satisfactory. What is the difference between these three samples and what is the impact on nucleic acid diagnostic experiments?
Simply put, plasma is part of whole blood and serum is part of plasma. Whole blood includes plasma and blood cells (erythrocytes, white blood cells, platelets), and plasma includes serum and fibrinogen.
From the perspective of nucleic acid diagnosis, human genomic DNA is more abundant in white blood cells, and only a small amount of free DNA is present in serum, but if white blood cells are largely apoptotic, the amount of free DNA in serum will increase. Pathogenic nucleic acids with pathogenicity are abundantly present in serum and are hardly present in blood cells.
Therefore, if human nucleic acid is used as a test target, such as screening for thalassemia, screening for color-blind genes, etc., whole blood should be used as an experimental sample because the amount of human nucleic acid contained in serum and plasma is much less than that of the whole. Blood, detection sensitivity will be much lower.
If the pathogen nucleic acid is used as the test target, such as detecting hepatitis B infection and detecting cold virus, etc., serum or plasma is used as an experimental sample, which naturally removes the blood cell impurity, which significantly reduces the difficulty in the nucleic acid extraction process. It is beneficial to improve the purity of nucleic acid and increase the sensitivity and stability of detection reaction.
Compared with serum and plasma, serum contains less fibrinogen. On the one hand, the DNA in serum is circulating DNA, which is mainly derived from diseased tissues. Therefore, using serum as an experimental sample can further improve the purity of the sample. Reduce protein impurities, increase the concentration of pathogens, and obtain more sensitive test results.
For the kit, the less sample impurities under the same system, the higher the quality of DNA extraction. Take the GNT series of blood DNA extraction kits as an example. If whole blood is used as a sample, the amount of blood added to each sample is usually 200 ul, and the yield is stable at 3-8 ug. However, if viral nucleic acid extraction is performed, serum should be taken as a sample. The amount of serum added to each sample should be 150~200ul. Compared with the blood kit system, the extraction efficiency of the serum kit will increase by 30% or more. Considering the purpose of the experiment, careful selection of the sample will result in better experimental results.
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