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文献和实验Optimal arbitrary primer length for Differential Display（图）
is determined by statistical consideration that each primer will recognize 50-100 mRNA species. To do so, these primers have to hybridize as 6-8mers (Liang and Pardee, Science. 1992, 257:967.). In practice, however, primers shorter than 9 bases failed to be used
from the primary RT-PCR, and using it as a template for a secondary round of PCR amplification. To avoid further amplification of primer-dimer artifacts or nonspecific products generated in the primary PCR, a different set of primers is employed in the secondary
Primer Dimer problems in real-time PCR-Real-Time PCR
. However, most of primers designed by Primer Express seem to have many dimer formation by upper or lower primers themselves or Upper-lower dimers. So what I can do is only choosing the set with smallest no, of dimer. So I have performed the normal PCR again and run gel
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