Eureka Genomics® Library Preparation Services for Next Generation Sequencing

Library Preparation of DNA or RNA for Next Generation Sequencing
Eureka Genomics performs library preparation of DNA or RNA for the purpose of next generation sequencing on Illumina's Genome Analyzer IIx. For convenience, the submitted DNA material can be isolated DNA, cDNA or amplified DNA. Low starting amount DNA will also be accepted for paired end, non indexed library preparation and sequencing. Submitted RNA material can be total RNA, mRNA or microRNA. Many sequencing and cycle choices are available. The library must be specifically prepared for the type of sequencing desired.

1. A Single End Non Indexed Library allows sequence data to be generated from only one end of the library construct. A Single End library is useful when the paired end information is not specifically needed. A Single End Library cannot be processed for paired end sequencing.

2. A Paired End Non Indexed Library allows sequence data to be generated from both ends of the library constructs. The paired end information can be used to distinguish structural rearrangements such as insertions, deletions and inversions. Paired end information can be useful in finishing genome assemblies. A Paired End Library can also be processed for single end sequencing.

3. Paired End Indexed Libraries can be used for two types of sequencing.

a. A Sample can be prepared as a Paired End Indexed Library for Multiplex sequencing. This is useful with samples that require paired end information but require a smaller amount of sequence data (<170MB). Up to 12 samples can be uniquely tagged with index sequences and can be combined into a single lane of a flowcell for Multiplex Sequencing. During data analysis the individual samples in the lane can be parsed out based upon their unique index tags.

b. Like the Paired End Non Indexed library, a Paired End Indexed Library can also be processed for single end multiplex sequencing while taking full advantage of the index. Single end multiplex sequencing allows sequence data from multiple samples to be generated in a single lane of a flow cell. This is useful with samples that require less sequence data and paired end information is not required. Depending on the amount of data required per sample, up to 12 samples can be uniquely tagged with index sequences and can be combined into a single lane of a flowcell for Multiplex Sequencing. During data analysis the individual samples in the lane can be parsed out based upon their unique index tags.

4. For mRNAseq a Paired End Non Indexed Library or a Paired End Indexed Library can be generated from mRNA (Items 2 and 3 above). The starting material can either be high quality total RNA or mRNA.

5. A Single End Non Indexed microRNA Library (Item 1 above) can be constructed from microRNA. The starting material can be either high quality Total RNA or pre-isolated microRNA.