一种改良的扩增cDNA5_末端的方法.pdf

2008, 35 ( 10): 1533- 1538 ActaH orticu lturae Sin ica cDNA 5 1, 3 2 1* 夏 瑞 , 陆旺金, 李建国 1 2 3 ( , 510642; , 510642; , 510640) : 5cDNA ( TdT ), TdT () PCR PCR PCR ( TaKaRa) , , ; , ; , : TdT; RACE; PCR; D l-Exp 1 : Q 9432 : A : 0513-353X (2008) 10-1533-06 A M odifiedM ethod of Amplifying the 5-end cDNA Sequence 1, 3 2 1* XIA Ru i , LU W ang-jin , and LI Jian-guo 1 2 ( China L itch iR esearch Cen ter, South China Agricultural University, Guangzhou 510642, China; Guang ong P rovincialK ey Laboratory of P ost-harvest S cience of F ru its an Vegetables, Guangzhou 510642, China; 3 Institute of F ruitResearch, Guang ong A ca emy of Agricultural Sciences, Guangzhou 510640, China) Abstract: In order to obtain the fu l-l length sequence of genesmore efficiently, amodifiedmethod of am- lifying the 5cDNA sequence, named asmodifiedTDT tailing am lifiedmethod, w as designed. M any strate- giesw ere used in this method includ ingTdT tailing, anchored PCR, nested PCR, touchdow n PCR. Com ared w ith a regu larmethod using a kit ( 5-Fu llRACE Core Set, TaKaRa), themod ifiedmethod hadmany advan- tages, such as smi le rinci le, easy o erability, h igh am lified s ecificity, and reliable results. Moreover, it costs less money and tmi e. Therefore, it isworth o ularizing in ord inary laboratories. K ey words: TdT; RACE; touchdown PCR; D l-Exp 1 , ( yeast two-hybrid) ( SSH ) ( genchi ) ( hagedis lay) , , (, 2005) , PCR PCR , , , PCR , 5 (, 2001) Frohman 1985 cDNA ( ra id am lification of cDNA ends, RACE) cDNA ( RACE) PCR, cDNA cDNA (, 2002), cDNA 3 5