
Thermo Reverse Transcriptase
- First-strand cDNA synthesis
- Active at 42~55℃. Optimum at 50℃.
- RNaseH negative
- Superior performance in RT-PCR. Up to 12 kb or more
- High processivity and sensitivity
Products
Cat.No. | Product | Size |
---|---|---|
RT10K | HelixCript™ Thermo Reverse Transcriptase | 10,000 units |
RT50K | HelixCript™ Thermo Reverse Transcriptase | 50,000 units |
RT10KN | HelixCript™ Thermo Reverse Transcriptase (with dNTP Mix) | 10,000 units |
RT50KN | HelixCript™ Thermo Reverse Transcriptase (with dNTP Mix) | 50,000 units |
- Description
- Data
- Details
- Documents
HelixCript™ Thermo Reverse Transcriptase, a Thermo stable and RNase H negative variant of M-MLV RTase, can synthesize cDNAs from purified RNA templates at a temperature range of 42℃ ~ 55℃ and especially shows the highest activity at 50℃. At the escalated temperature, the cDNA synthesis is enhanced partially due to the less internal structural formation of the template RNA and increased polymerization activity of RTase. The high processivity and productivity of HelixCript™ Thermo Reverse Transcriptase allows this enzyme to amplify the high yield of product and can synthesize cDNA of target gene up to 12 kb from RNA template. This enzyme is quite suitable to synthesize the first-strand cDNA for RT-PCR of target gene. The cDNA synthesis from total RNA or poly-(A)+ RNA is performed by random primer, oligo-d(T) primer, or gene-specific primer.
ebiomall.com






>
>
>
>
>
>
>
>
>
>
>
>
二、限制性内切酶消化样品DNA
三、凝胶电泳分离消化产物
四、如果靶序列>5kb,在0.25M的HCl中进行震荡脱嘌呤10min,ddH2O洗一次
五、用变性液震荡处理30min,ddH2O洗一次
六、中和液震荡处理30min
七、裁取合适大小的尼龙膜或硝酸纤维素膜进行转膜操作,可用真空转膜仪或者搭滤纸桥,需要20×SSC
八、制备探针,可用地高辛标记(以地高辛为例)
九、将膜放入杂交瓶,42°C预杂交30min
十、倒掉预杂交液,加入杂交液,适当的温度进行杂交4h至过夜
十一、洗膜,先用2×SSC+0.1%SDS,20-25°C洗2×5min,再用0.5×SSC+0.1%SDS洗2×15min。然后用washing buffer洗1-5min,接着用blocking solution洗30min后,用antibody solution洗30min,再用washing buffer洗2×15min,再用detection buffer洗2-5min后,取出膜,放于保鲜膜上,在结合有DNA的一面滴加CSPD ready-to-use后,立刻盖上保鲜膜,让CSPD ready-to-use均匀的布满膜表面,室温放置5min后,37°C温育10min以上
十二、放射自显影,可用成像系统信号累积模式显影或用X-ray显影

