![]() ![]() The sequence of the gene is from 5’ to 3’ direction. It starts from the start codon (ATG) and ends with the stop codon (TAA). We copied the Dsup gene nucleotide sequence from the NCBI portal (mentioned above) and pasted it in the APE tool.The primer design steps remain the same even if you use a different tool.įollowing are the steps involved in designing forward primer: We are using APE (A Plasmid Editor) software to design the primers, which is free to use. To follow along, access the sequence of Dsup from this link. The following content focuses on demonstrating designing primers for different cloning methods. In case, if you take the complementary sequence and use it for primer designing (laying the sequence in 5’-3’), the reverse primer in the above example now acts as the forward primer. The forward primer creates copies of the 5’-3’ strand whereas the reverse primer makes copies of the complementary (runs 3’-5’) strand. Whereas Reverse primer uses the upper strand as a template and synthesizes the lower strand.Īlthough the names suggest they create copies of different strands, their names depend on the direction of the strand being used for amplification. ![]() The forward primer synthesizes the upper strand using the bottom strand as a template. One is called ‘forward primer' and the other one is called ‘reverse primer’. To amplify any DNA sequence, two primers are necessary. Dsup protein is a newly discovered gene that imparts resistance to UV radiation by coating itself to DNA. Tardigrades are fascinating animals with extraordinary abilities to cope with extreme conditions like the vacuum of space, high tolerance for UV radiation, high and low-temperature tolerance to begin with. The primer design is demonstrated using Dsup (Damage Suppressor) gene from tardigrade (water bear). This article demonstrates how to design primers (forward and reverse) for different types of cloning methods. ![]()
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