玉米免DNA制备单株SSR和SCAR基因分型方法研究

A fast SSR and SCAR genotyping method without DNA preparation in maize

探讨采用免DNA制备法分析玉米单株SSR与SCAR分子标记的基因型。分别刮取各单株少许叶肉细胞到1×PCR缓冲液中,置于PCR仪上95℃10 min,冷却,添加PCR其它组份,完成SSR与SCAR的PCR扩增与显色成像。免除模板DNA制备过程,对掖478与丹340两个亲本及57个(共58个)F2分离群体单株分别实现SSR标记-phi402893基因型快速鉴定,对掖478与丹340两个亲本及5个F2分离群体单株分别实现SCAR标记-MITE185基因型快速鉴定。

The objective of this study is to exploit the possibility of SSR and SCAR genotyping of maize individual plant without DNA preparation. Several cells were scratched and put into 1×PCR buffer,boiled at 95℃ for 10 min in PCR thermocycler and then cooled down. The other PCR reaction components were added and SSR ＆ SCAR PCR amplifications were finished. SSR locus, phi402893, had been genotyped successfully among Ye478 and Dan340 along with 57 individuals of their F2 progeny （58 in totals） as described method. MITE185,one of SCAR markers of Dwarf8 gene associated with early maturity, had been genotyped successfully among Ye478 and Dan340 along with 5 individuals of their F2 progeny as described method. The results suggest that the rapid SSR and SCAR genotyping without DNA preparation could be used into molecular marker assisted selecting the target genotypes among individuals.