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It says that i have to pick primers between 17-21 bp which are found in the last half of the cDNA. How come the last part of the cDNA?
Thanks.
Thanks.
"Selecting Primers 17-21 bp from cDNA Last Half" refers to the process of choosing short segments of nucleotides (building blocks of DNA) that will be used to initiate the amplification of a specific section of cDNA (complementary DNA) in a laboratory setting.
Selecting the right primers is crucial because they determine the specificity and efficiency of the amplification process. If the primers do not match the target cDNA sequence accurately, the desired amplification will not occur. Additionally, using primers that are too long or too short can result in non-specific amplification or low amplification efficiency, respectively.
The recommended length for primers in cDNA amplification is between 17-21 base pairs (bp). This length is considered optimal for ensuring specificity and efficiency of the amplification process, as well as minimizing the likelihood of primer-dimer formation.
To select the best primers for cDNA amplification, one should first identify the target cDNA sequence and then use specialized software or online tools to design primers that meet the recommended length and other criteria such as melting temperature, GC content, and absence of hairpin structures. It is also important to validate the primers through experimental testing.
Some common challenges in selecting primers for cDNA amplification include finding primers that are specific to the target sequence, avoiding primer-dimer formation, and minimizing the presence of secondary structures. Additionally, the presence of repetitive sequences or genetic variations in the target cDNA can also pose challenges in primer selection.