mRNA Sample Preparation

The LNA™-enhanced oligo-T20 capture probe of our mRNA Sample Preparation kit results in efficient isolation of mRNA directly from guanidinium thiocyanate-containing cell or tissue extracts.

Ina K. Dahlsveen, Ph.D., Product Manager

High yield mRNA isolation directly from guanidinium thiocyanate-containing cell or tissue extracts
mRNA isolation without the influence of endogenous RNases
50-fold higher recovery of mRNA than obtained using DNA oligo-dT for mRNA capture
Compatible with beads, tubes and microtiter plates
Fully developed protocols

Features

This product is designed for efficient isolation of polyadenylated mRNA directly from lysed guanidinium thiocyanate (GuSCN)-containing cell or tissue extracts. This is achieved by combining the use of a biotinylated LNA™ oligo(T) capture probe with subsequent immobilization of the captured poly(A)+RNA.

Coverage

The LNA™-enhanced oligo-T20 capture probe is available in two formats:

5’-biotinylated for capture on streptavidin-coated magnetic particles
5’-amino-labeled for capture onto solid surfaces

Fully developed protocols for mRNA isolation from various sample types are available (see right-hand menu)

Unlike DNA oligonucleotides, LNA™ oligonucleotides can form duplexes with their complementary sequences in strong chaotropic buffers even at elevated temperatures. LNA™ oligonucleotides can therefore be used for poly(A)+RNA selection directly from guanidinium thiocyanate (GuSCN) cell or tissue extracts under the most stringent conditions.

This means that mRNA can be captured under conditions where nucleases and proteases are inactivated and where proteins are dissociated from the target mRNA, thereby providing optimal access to the target sequence.

The LNA™-enhanced oligo-T20 capture probe from Exiqon recovers up to 50 times more mRNA from GuSCN cell extracts than DNA oligo-dT capture probes.

Figure 1

GuSCN concentration on the recovery of yeast THI4 mRNA

LNA™ captures mRNA more efficiently than DNA. (Click to learn more)

Figure 2
recovery (%) of the yeast THI4 mRNA by the DNA-T20 probe and the LNA™-

LNA™ probes are more efficient at high GuSCN concentrations than DNA probes. (Click to learn more)

Figure 3
recovery (%) of polyadenylated yeast ACT1 spike mRNA in 4 M guanidinium thiocyanate