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Storage and Stability

Store all system components at -20℃ for one year.

Description

The TonkBio^TM First Strand cDNA Synthesis Kit for RT-PCR is optimized to efficient synthesize first strand cDNA from mRNA or total RNA templates. The kit uses M-MLV reverse transcriptase with lower RNase H activity, which avoids degradation of RNA when RNA-DNA hybrid forms in the progress of cDNA synthesis. It is suitable for synthesis of cDNA up to 9 kb.

The cDNA products could be used directly in a variety of downstream applications, including PCR, qPCR, gene clone.

Caution

1. For research use only.

2. Use RNase-free reagents during RNA extraction and cDNA synthesis process.

3. NO repeated freezing and thawing RNA. RNA should be dissolved in ice-water mixture.

4. The reverse transcription reaction products should be stored at -20℃.

5. Wear gloves when handling RNA and all reagents as skin is a common source of RNases.

Example of application

Troubleshooting Guide

1. NO or low product of RT PCR

1）Degradation of RNA template. RNA Purity and integrity is important to obtain full length cDNA. Users have to make gel electrophoresis detection to ensure no degradation occurred. And avoid repeated freezing and thawing RNA.

2）Low concentration of RNA template. Trace amount of agents used in RNA purification may remain in solution and inhibit cDNA synthesis. Suggest to wash RNA with 75% ethanol ( prepared by DEPC water) carefully.

3）Incorrect primers. Select random primers or specific primers instead of OligodT primer with bacterial RNA or RNA without a poly A tail.

4）GC rich template. If the RNA template is GC-rich or contains secondary structures, choose random primers and increase the reaction temperature of transcription up to 55℃.

2. RT-PCR product longer than expected

RNA sample is contaminated with DNA. To avoid genomic DNA contamination, User could use DNase I to digest RNA sample before transcription or design primers on exon-intron boundaries.

3. Product of RT-PCR in negative control

RNA template is contaminated with DNA. Perform DNaseI digestion prior reverse transcription.

免费试用说明：

1.运费：申请机构（单位）无需承担试用装的运输费用

2.发货时间：申请审核通过后，协商发货时间；

3.申请要求：申请实验室有相应实验，能够近期开展实验；

4.其它说明：请提供试用报告，谢谢配合！