利用switching mechanism at 5′end of RNA transcript(SMART)技术对猪单个体外成熟培养的MⅡ期卵母细胞进行了扩增,在此基础上,利用T7体外转录技术进行体外转录,aRNA浓度达到1.962μg。通过联合SMART技术和T7体外转录技术,mRNA浓度扩增了2.3×104倍左右,这个数量级已基本能满足cDNA array等通量筛选研究的需求。本研究为珍贵和微量的生物材料进行高通量筛选研究提供了一个有效的体系。
Abstract:
By means of switching mechanism at 5' end of RNA transcript(SMART)technology,single matured porcine MⅡ oocyte cDNAs were amplified.Meanwhile,the mRNA was amplified from the single porcine oocyte cDNA products using T7 RNA polymerase based in vitro transcription technology(IVT),and the aRNA concentration reached 1.962 μg.Smart technology combined with IVT technology,mRNA has augmented 2.3×104 times,and this amount could meet the cDNA array and other high throughput screening requirements.Based on this research,a feasible platform has been established to carry out high throughput screening researches for precious and rare biological materials.
