Rapid and automated isolation of mRNA

Dynal Biotech's new range of Dynabeads mRNA isolationproducts enable even the most difficult sources - includingembryos - to be used to provide material suitable for downstreamHTS gene expression studies.

"The reliability and simplicityof Dynabeads separation makes them the mRNA isolation method ofchoice for gene expression," says Tone Yrvum, productmanager, molecular systems.

"Separations are easilyscaleable; mRNA can be isolated from any quantity of cells andcan be used to prepare mRNA for any type of microarray,macroarray, or chip.

"Whatever the starting material andintended destination, Dynabeads will get you from A to B quicker,easier and more consistently than any other protocol."Actually, the separation of mRNA takes only 15minutes".

Based on superparamagnetic separationtechnologies, pioneered and developed by Dynal Biotech, DynabeadsOligo (dT)25, and associated kits, can consistently isolate polyA+ mRNA with or without prior total RNA preparation, it isclaimed.

More importantly, the source of RNA can be controlledusing Dynal Biotech's cell-specific Dynabeads to selectcells based on distinctive surface marker proteins.

Thesemagnetic separations avoid the complications of centrifugation orother cell sorting procedures, thus decreasing degradation byreducing the time between cell/tissue harvesting and RNAextraction.

The beads themselves are highly engineered to provideuniform spheres with even magnetic properties and maximum surfacearea, and remain dispersed in solution.

Their chemically inertnature allows downstream applications to be performed whilst themRNA is immobilised on the bead, producing a reusable platformfor gene-expression studies - evidence also shows that RT-PCR ismore efficient.

The beads and mRNA can be recovered at variousstages of a reaction and can therefore be used to produce astable, reusable cDNA library - especially useful when originaltemplate is at a premium.

The magnetic separation process notonly supports numerous downstream applications, but also ensuresthat they are free from contaminating material, since the processdoes not use chemicals classically associated with RNA extractionsuch as DTT, chaotropic salts, and DNase.

Instead, with the RNAbound to the beads, the unwanted material remains in solution andis decanted.

The ease of this isolation procedure facilitates theuse of liquid handling platforms, such as Tecan's FreedomEvo or Beckman Coulter's Biomek FX, to greatly increasesample throughput.

Dynal Biotech's mRNA Dynabeads kitsinclude Dynabeads mRNA Purification kit for isolation from totalRNA; Dynabeads mRNA Direct for isolation direct from cell andtissue lysates; Dynabeads mRNA Direct Micro for isolation fromcell and tissue lysate with subsequent reusable cDNA templateproduction for RT-PCR; Dynabeads Leucocytes mRNA and MonocytesmRNA isolation kits specifically for whole blood leucocyte andmonocyte mRNA isolation respectively; and Dynabeads oligo (dT)25for scalable standard and de novo applications.