Abstract:
:We report a rapid and simple method for isolating the 5'-end of plant genes from genomic DNA by polymerase chain reaction (PCR) with TATA-box-based degenerate primers (TDPs). The TDPs were specially designed according to the TATA box, which is conserved in the promoter region of most plant genes. The unknown 5'-ends of several genes in different plants were isolated by PCR with gene-specific primers of the known core fragment and the TDPs. Our method does not require the arduous RNA manipulations and expensive enzyme treatments of the popular rapid amplification of cDNA ends (RACE) and its variants, and so could be a cheap practical alternative.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Guo Y,Ma L,Ji Y,Pu G,Liu B,Du Z,Li G,Ye H,Wang Hdoi
10.1007/s12033-010-9323-0subject
Has Abstractpub_date
2011-02-01 00:00:00pages
152-6issue
2eissn
1073-6085issn
1559-0305journal_volume
47pub_type
杂志文章abstract::Tea, a beverage crop, is a rich source of polyphenols and polysaccharides which greatly attribute to its importance. However, oxidation and precipitation of these compounds during nucleic acids extraction is a limitation to molecular biology and genomic studies. On isolation of total RNA from root tissue using establi...
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