The amplicon library was sent to emPCR amplification and 454 sequencing as described previously [25, 26. Briefly, LibL emPCR Kit (Roche) was used for emPCR according to the emPCR Amplification Method Manual. Accordingly, singlestranded template and emulsion PCRs were performed according to the emPCR Amplification Method ManualLibA (Roche Applied Science). DNA sequencing was performed in a GS Junior NGS platform. Mutation analysis with random DNA identifiers (MARDI) catalogs Piga mutations in heterogeneous pools of CD48deficient T cells derived from DMBAtreated rats.
Authors. Javier R. Revollo, Corresponding author. (emPCR Amplification Method Manual LibL for GS Junior). The resulting beads were washed, screened for DNAcoating, combined with A library titration was carried out by doing an emulsion PCR in smallscale (SVemPCR), using the GS Titanium SVemPCR Kit LibL v2 (Roche) and following the protocol detailed in emPCR Amplification Method Manual Lib L SV (May 2011). Emulsion PCR was performed according to the manufacturers instructions (emPCR Amplification Method ManualLibL, GS Junior Titanium Series, March 2012).
To perform the emPCR an input of 0. 7 molecules per bead was chosen. FullText Paper (PDF): Systematic Comparison of Three Methods for Fragmentation of LongRange PCR Products for Next Generation Sequencing Whole genome and normalized mRNA sequencing reveal genetic status of TK6, WTK1, and NH32 human Blymphoblastoid cell lines The emPCR was performed according to the emPCR Amplification Method ManualLibL GS Junior Titanium Series (May 2010; Rev.
April 2011) Location of variations in mRNA. EmulsionPCR was carried out with GS Junior Titanium emPCR Kit (LibL, 454 Life Sciences Corp.CT, USA) 6 according to the emPCR Amplification Method Manual LibL, SinglePrep. The sequencing reaction was carried out with the GS Junior Titanium Sequencing Kit and GS Junior Titanium PicoTiterPlate Kit (454 Life Sciences Corp.
) according to The gray bars above the graph depict the location and length PCR fragments [in kbp and the red squares highlight the seven coding exons of the ENST transcript. annealed to capture beads, and clonally amplified by emulsion PCR (emPCR, Amplification Method Manual LibL; GS Junior Titanium Series, Roche). After emPCR, beads with