Mint and Mint-Universal cDNA synthesis kits

cat.# SK001; SK002

- Fast cDNA synthesis protocol
- High content of full length transcripts
- Low background
- Free trial-size Encyclo PCR kit included

Mint and Mint-Universal cDNA synthesis kits are designed to synthesize full-length-enriched double stranded (ds) cDNA from total or poly(A)+ RNA. cDNA prepared by these kits are flanked by known adapter sequences at the both ends and can be used for various applications including construction of cDNA libraries, virtual Northern blot, subtractive hybridization, array probe preparation.

Method overview

MINT cDNA synthesis is based on a novel technology (patent pending) utilizing the specific features of MMLV-based reverse transcriptase (RT). First strand cDNA synthesis starts from the 3'-primer comprising an oligo(dT) sequence to anneal to the poly(A)+ stretch of RNA. When RT reaches the 5' end of the mRNA, it adds several non-template nucleotides, primarily deoxycytidines, to the 3' end of the newly synthesized first-strand cDNA (Schmidt and Mueller, 1999). This oligo(dC) stretch base pairs to the complementary oligo(dG) sequence located at the 3' end of a special 30-mer deoxyribooligonucleotide called PlugOligo. RT identifies PlugOligo as an extra part of the RNA-template and continues first strand cDNA synthesis to the end of the oligonucleotide, thus incorporating PlugOligo sequence into the 5' end of cDNA.

The last 3'-dG residue of the PlugOligo is a terminator nucleotide comprising 3'-phosphate group. This blocking group prevents unwanted annealing and extension of the PlugOligo. Under standard conditions RT can hardly use PlugOligo as a template, however our special IP-solution (solution for Incorporation of PlugOligo sequence) dramatically increases the efficiency of this process. Synthesis of ds cDNA is then performed using PCR amplification.

	Schematic outline of Mint cDNA synthesis. Enlarge scheme

Mint-amplified cDNA from different sources. 1 - mouse liver; 2 - mouse skeletal muscle; 3 - mouse brain; 4 - human leucocytes; 5 - human lung; 6 - human skeletal muscle; 7 - mosquito grub; 8 - copepod Pontella sp.; 9 - tomato Lycopersicon esculentum. M - 1 kb DNA size marker, SibEnzyme, Russia.