引物问题（Primer problem）.docVIP

引物问题（Primer problem） What are the basic principles of 30. primer design? The following principles of primer design are for your reference: 1) primers are best designed in the conserved region of template cDNA. 2) the length of primers is usually between 15-30 bases. 3) the content of primer GC is between 40%-60%, and the Tm value is best to be close to 72 centigrade. 4) the primer 3 end should avoid codon third. 5) the primer 3 end cannot select A, and the best choice is T. 6) the base is randomly distributed. 7) there should be no complementary sequence between primers and primers. 8) primer 5 end and the middle G value should be relatively high, and the 3 end of a low G value. 9) the 5 end of the primer can be modified, but the 3 end is not modified. 10) single strand of amplification product can not form two stage structure. 11) primers should be specific. 31. what are the design software of common primers? Commonly used software has Oligo 6 and Primer Premier 5. Primer design software is based on the design guidelines for primer design. In fact, the key to the success of PCR amplification is the preparation of reaction templates and the control of reaction conditions. The disadvantage of primer design software is that sometimes it is judged that no part of the gene meets the requirements of standard primers. Genscript provides you with the following primer design software: Primer calculation tool Primer design tool Sequence primer design software Real-time PCR primer design software 32. can the primers and probe sequences found in the literature be used directly? Foreign literature credibility is usually relatively high, can be used directly; but for insurance purposes, the best use of the blast sequences of the primer probe to carry out the necessary verification; or further analyzed by primer design software primer probe of two grade structure and annealing temperature, which is more conducive to your understanding of the whole experiment. 33. how do we calcul