A primer is a short nucleic acid sequence that provides a starting point for DNA synthesis. In organisms, primers are short strands of RNA. Before DNA replication can occur, primers must be synthesized by an enzyme called primerase, which is an RNA polymerase. The synthesis of primers is necessary because the enzyme that synthesizes DNA (called DNA polymerase) can only connect new DNA nucleotides to existing nucleotide strands. Therefore, primers are used to prepare and lay the foundation for DNA synthesis. Remove primers before DNA replication is complete, and use DNA polymerase to fill in the gaps in the sequence with DNA. In the laboratory, scientists can design and synthesize DNA primers with specific sequences that bind to the sequences of single-stranded DNA molecules. These DNA primers are commonly used to perform polymerase chain reactions to replicate DNA fragments or to sequence DNA. Oligonucleotide primers are required for PCR reactions. It is necessary to design primers that are complementary to the DNA template region. They are chemically synthesized by linking nucleotides. When adding one nucleotide at a time, the reactive group on the nucleotide must be selectively blocked and repeatedly unblocked. The main characteristic of primers is that they must correspond to the sequence of the template molecule (they must be complementary to the template strand). However, the primer does not need to match the template strand exactly; however, the 3'end of the primer must match the template DNA strand exactly in order to continue extension. Guanine or cytosine is usually used at the 3'end, and the 5'end of the primer usually has a few nucleotide extensions. Also, the two 3 'ends of the hybrid primer must point towards each other. The size of the primer is also important. Short primers are used primarily to amplify small, single DNA fragments. On the other hand, long primers are used to amplify eukaryotic genomic DNA samples. However, the primer should not be too long (> 30 primer units) or too short. Short primers will produce inaccurate and non-specific DNA amplification products, while long primers will reduce the rate of hybridization. On average, the size of the DNA fragment to be amplified should be within 110 kB.

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