T3 酶混合物、10X 反应缓冲液、2XNTP/CAP 溶液、GTP 溶液、pTRI-Xef、TURBO DNase、醋酸铵终止液、氯化锂沉淀溶液和凝胶上样缓冲液 II 均储存在 –20°C 下。无核酸酶水可在任何温度下储存。
常见问题解答 (FAQ)
I am using the mMESSAGE mMACHINE T3 Transcription Kit for in vitro transcription. I ran my reaction product on a denaturing gel and got a smear. Can you offer some advice?
If the RNA appears degraded (e.g. smeared), remove residual RNase from the DNA template preparation before in vitro transcription. Do this by digesting the DNA prep with proteinase K (100-200 µg/mL) in the presence of 0.5% SDS for30 min at 50°C, follow this with phenol/chloroform extraction. The RNase Inhibitor that is present in the transcription reaction, can only inactivate trace RNase contamination. Large amounts of
RNase contamination will compromise the size and amount of transcription products.