cDNA Synthesis Kit

M-MLV cDNA Synthesis kit provides a sensitive and easy-to-use solution for two-step RT-PCR. This kit includes five tubes comprehensive of the reagents required for successful RT-PCR. The M-MLV H-reverse transcriptase is optimized for reliable cDNA synthesis over a wide dynamic range of input RNA. The enzyme is exceptionally well with a wide variety of targets.

long time at -20°C

1. M-MLV RTase H-
2. 5x RT buffer (contain dNTP mix)
3. 5x RNA protector
4. oligodT (15mer)
5. Random primer (6mer)

1. First-strand cDNA synthesis for subsequent PCR or real-time PCR.
2. RT-PCR validation of gene expression data obtained from microarray experiments.
3. RT-PCR validation and quantification of gene silencing by RNA interference.

200 units of enzyme are used to produce cDNA from 1µg of a 1.2kb control RNA. The minimum specification is 120ng of first strand cDNA made from 1µg of RNA. The cDNA product must be >90% full length.

To test for endonuclease activity, 1µg of Type I supercoiled plasmid DNA is incubated with 500 units of M-MLV Reverse Transcriptase in 1X Reaction Buffer for one hour at 37° C. Following incubation, the supercoiled DNA is visualized on an ethidium bromide-stained agarose gel to verify the absence of visible nicking or cutting (analysis on 0.4µg of DNA).

The purity is >90% as judged by SDS-polyacrylamide gels with Coomassie® blue staining.