How to extract dna from blood clot
Hello,I am trying to extract DNA from clotted blood and I found some protocols online that suggest treating the blood with glycogen. The glycogen is suppose to improve the yield. I was wondering if anyone could tell me specifically what the glycogen does. I am studying for clinical licensure and this is what it says to do in my study guide if you get a blood sample with clots. AbstractThe clot from blood is usually discarded after the collection of serum.
Yet, it contains nucleated white blood cells and substantial serum. Here, we have compared four methods to enable quick and efficient extraction of human genomic and viral DNA from clotted blood. Two of these methods, a phenol-based in-house method and Tripure isolation reagent, only achieved a low polymerase chain reaction (PCR) yield. In contrast, the QIAamp blood kit and the High Pure Viral Nucleic Acid kit were equally MethodsHigh-speed shaking of clot samples in a cell disruptor manufactured for homogenization of tissue and faecal specimens was evaluated for processing frozen blood clots for DNA extraction.
The method was compared to two commercial clot protocols based on a chemical kit and centrifugation through a plastic sieve, followed by the same DNA extraction protocol. ResultsHigh speed shaking succeeded in fully dispersing the clots and the method generated the highest DNA yield. The level of PCR detection of P. falciparThe blood clot s DNA of human beings was extracted respectively by three methods such as higher sodium chloride method,silica gel method and improved phenol-chloroform method,and the degree of purity and the content of the DNA were determined by ultraviolet photometer and agarose gel electrophoresis separately.The results showed that the DNA extraction time of silica gel method was the shortest,and the content of DNA was higher,meanwhile the method of improved phenol-chloroform was the easiest,but the pollution was serious by using organic solvent, whereas higher sodium chlorid method needed a great amount of blood clots and the content of DNA was not stable.