Libraries are prepared from mRNA by construction of a cDNA library followed by preparation of a DNA library using the quick library preparation method for the 454™ sequencing platform. This method includes repair of DNA ends and the addition of a non-templated dAMP. Libraries can also be constructed using the original 454 library construction method. However, this method is no longer commonly used. For more information, choose the workflow tab below.
Libraries prepared in this manner are suitable for capture bead immobilization and emulsion PCR, the next steps in the 454 sequencing workflow.
NEBNext® reagents are available for each step in the mRNA library preparation workflow for 454 sequencing. Reagents from NEB are available in a set, master mix or module format. Each reagent has been functionally validated by preparation of an RNA library from a standard reference followed by 454 sequencing.
454™ is a trademark of Roche.
NEBNext® is a registered trademark of New England Biolabs, Inc.