Villalva C, Touriol C, Seurat P, Trempat P, Delsol G, Brousset P
Centre Hospitalier Universitaire de Purpan Toulouse, France.
Biotechniques. 2001 Jul;31(1):81-3, 86. doi: 10.2144/01311st04.
Under certain conditions, T4 gene 32 protein is known to increase the efficiency of different enzymes, such as Taq DNA polymerase, reverse transcriptase, and telomerase. In this study, we compared the efficiency of the SMART PCR cDNA synthesis kit with and without the T4 gene 32 protein. The use of this cDNA synthesis procedure, in combination with T4 gene 32 protein, increases the yield of RT-PCR products from approximately 90% to 150%. This effect is even observed for long mRNA templates and low concentrations of total RNA (25 ng). Therefore, we suggest the addition of T4 gene 32 protein in the RT-PCR mixture to increase the efficiency of cDNA synthesis, particularly in cases when low amounts of tissue are used.
在某些条件下,已知T4基因32蛋白可提高不同酶的效率,如Taq DNA聚合酶、逆转录酶和端粒酶。在本研究中,我们比较了添加和不添加T4基因32蛋白时SMART PCR cDNA合成试剂盒的效率。将该cDNA合成程序与T4基因32蛋白结合使用,可使RT-PCR产物的产量从约90%提高到150%。对于长mRNA模板和低浓度的总RNA(25 ng),甚至也能观察到这种效果。因此,我们建议在RT-PCR混合物中添加T4基因32蛋白,以提高cDNA合成的效率,特别是在使用少量组织的情况下。