AD Sequencing MethodsThey have made great strides in recent years, solving historical crimes and ancient mysteries. Or even a combination of both. That’s what a team of geneticists has just done. Turi Kingbelonging University of LeicesterThe researcher is known for being behind the DNA sequencing that helped identify King Richard III in 2014, so they had no hesitation in counting on her finally trying to solve the mystery. historical mystery is considered one of the most mysterious of the 19th century.
Again, it’s something to do with aristocracy. In particular, the personality Kaspar Hauser“wild child” who was brutally murdered after being suspected of being the missing son of a prince from the German region of Baden.
After his murder, fragments of his bloody clothing were preserved. So when DNA sequencing techniques began to be used more widely in the 1990s, several attempts were made to compare him with those of his supposed sisters to find out whether he really was the lost heir. Unfortunately, the DNA was too damaged and sequencing techniques were still in their infancy, so it was impossible to find out whether Hauser was the Little Prince. Now, 30 years later, the DNA is still just as damaged or even more, but a great revolution in genetics has finally provided a solution to this mystery.
A history of deaths and betrayals
In 1812, the only male child Grand Duke CharlesPrince of Baden, died in infancy. As a result, the prince was left without an heir and a long dispute began over the creation of alternative lines of succession.
The dispute was still in the spotlight when, 16 years later, the city Nuremberg with two letters in which he stated that he had grown up in a dungeon, imprisoned by a mysterious man. He could barely speak and certainly could not write, but he had been taught to write his name: Kaspar Hauser.
The boy was teenagerabout 16 years old, so he was suspected of being the son of the Grand Duke. At the time, after the infant’s death, it was thought that the prince’s enemies might have kidnapped his heir, leaving the dying infant in his crib to make it appear that he had died. This would have facilitated the desired alternative line of succession.
However, it was never proven whether this had actually happened. Hauser’s appearance added fuel to the fire of this theory, so from the shadows, opponents of the legitimate inheritance of the Grand Duke’s son put a price on his head. Five years later, the poor young man was stabbed to death. But it was never known whether it was really the prince or the work of a fraud. Thus was born this historical mystery, which was finally solved thanks to DNA sequencing methods.
A Brief But Remarkable History of DNA Sequencing
In the 1977s, two main methods of DNA sequencing came into use. dideoxynucleotide methoddeveloped by Sanger, and chemical methoddeveloped by Maxam and Gilbert. Both are based on the fact that DNA is made up of nucleotides linked to four different types of nitrogenous bases: adenine (A), guanine (G), cytosine (C), and thymine (T).
The Sanger method mimics the natural method of DNA replication using an enzyme called DNA polymerasewhich takes a strand of DNA and connects each nucleotide to another nucleotide using a complementary base. Adenine and thymine are complementary to each other, as are guanine and cytosine. The difference between the natural method and Sanger DNA sequencing is that in each cycle, a dideoxynucleotide is added for a particular base.
These are nucleotides that absence of hydroxyl groupconsisting of hydrogen and oxygen and stopping the process when it places. That is, the polymerase does not continue to place nucleotides. In this way, it is possible to measure the fragments formed in each cycle and distinguish where the various bases that make up the letters of DNA are located.
Chemical sequencing uses substances that create breaks in different bases. Thus, later the sequence composition can also be made by analyzing the resulting fragments.
This is all a very slow process. However, over the years it has been automated by machines that have become much better at analyzing sequences. This made it possible to begin sequencing the human genome in the 1990s, opening up a huge range of possibilities. You can look for genes associated with human diseases, identify a person’s paternity, or even solve a historical mystery, like Hauser. Unfortunately, there was still disability.
A historical mystery that is difficult to solve
In the 1990s and 2000s, genetic sequencing techniques could only analyze large fragments of DNA. Unfortunately, DNA Over the years it deteriorates.therefore, only short fragments can be obtained from old samples. For this reason, Hauser’s first attempts to sequence DNA were unsuccessful.
The samples were stored until King’s team decided to reopen the investigation. Using new DNA sequencing techniques, they were able to solve this historical mystery once and for all.
To do this, they focused on mitochondrial DNA. It is found in the mitochondria, the organelles that serve as the engine of the cell, as this is where cellular respiration occurs. It is DNA that is primarily passed down through the maternal line and is very interesting for analyzing ancient samples, as it can be found in large quantities and is not easily lost through degradation.
King’s team began by analyzing mitochondrial DNA from blood samples from Hauser’s clothing, as well as from hair taken from a corpse. The DNA was proven to be identical, so the samples were genuine. However, what was unexpected was that they decided to compare it with the DNA of the Grand Duke’s other daughters, as well as their descendants. The samples showed no connection.
This shows that poor Kaspar, who was killed for being the son of the Prince of Baden, was not even that person. He may have been the double victim of murder and fraud. Not to mention the terrible conditions he had to grow up in until he was 16 years old, barely able to speak. Unfortunately, this will remain a mystery until the end of history.
Source: Hiper Textual
