100bp paired-end RNA-seq library

[miu007]

Hi folks,

Does anyone have experience making libraries for 100bp paired-end sequencing? What is the optimal insert size? How to generate fragments suitable for this purpose?Thanks.

[luc]

These two protocols are very good:

High-Throughput Illumina Strand-Specific RNA Sequencing Library Preparation.
Cold Spring Harb Protoc. 2011;2011(8)
Authors: Zhong S, Joung JG, Zheng Y, Chen YR, Liu B, Shao Y, Xiang JZ, Fei Z, Giovannoni JJ


A Low-Cost Library Construction Protocol and Data Analysis Pipeline for Illumina-Based Strand-Specific Multiplex RNA-Seq
Lin Wang, Yaqing Si, Lauren K. Dedow, et al. 2011 PLoS ONE 6(10)