Deev S M, Barbakar' N I, Karlyshev A V, Sakharova N K, Grechko V V
Mol Biol (Mosk). 1980 Mar-Apr;14(2):413-20.
cDNA was synthesized on Ig kappa-L-chain mRNA isolated from mouse plasmocytoma MOPC 21 using avial myeloblastosis RNA-dependent DNA polymerase. The cDNA was used as a matrix for second DNA chain synthesis using 5'-exonuclease free DNA-polymerase I (Klenoff fragment). Hairpin DNA having double length of initial cDNA was obtained without added exogenous primer. SI nuclease treatment of the hairpin DNA results in reducing the DNA length twice, as shown by electrophoresis in denaturing conditions. The fact, that S1 nuclease resistance of the hairpin DNA is 75% shows high complementarity between first and second DNA chain. The product length obtained after S1 nuclease treatment was shown to be heterogenous. The maximal length of the product, reaching at least 900 base pairs, is near to be initial mRNA length (without the poly(A)-fragment).
使用禽成髓细胞瘤RNA依赖性DNA聚合酶，从小鼠浆细胞瘤MOPC 21分离的Igκ轻链mRNA上合成cDNA。使用5'-外切核酸酶-free DNA聚合酶I（Klenow片段），将该cDNA用作第二DNA链合成的模板。在不添加外源引物的情况下，获得了具有初始cDNA两倍长度的发夹DNA。如变性条件下的电泳所示，对发夹DNA进行S1核酸酶处理会使DNA长度减半。发夹DNA对S1核酸酶的抗性为75%，这一事实表明第一和第二DNA链之间具有高度互补性。S1核酸酶处理后获得的产物长度显示为异质性。产物的最大长度至少达到900个碱基对，接近初始mRNA长度（不包括poly(A)片段）。
