Next Generation Sequencing Request Form
Sample submission details
For Sanger sequencing, fragment analysis and user-prepared NGS pools the sample submission information is detailed on the second sheet of the request form.
Total RNA for RNA sequencing NGS library preparation
Total RNA submitted for NGS RNA-seq (using either mRNA isolation or rRNA depletion) must be free from contaminating DNA, salts (e.g. Mg2+, or guanidinium salts, divalent cation chelating agents (e.g. EDTA or EGTA) or organics (e.g. phenol or ethanol). We do not recommend specific RNA isolation kit but column, or magnetic-bead based methods, which include a DNase digestion step, are known to provide RNA of sufficient purity. RNA isolated using organic methods (e.g. TriZol reagent) should be cleaned-up, and DNase treated, with a column based kit. Please elute your RNA in nuclease free water.
If it is not possible to provide high purity RNA please contact us to discuss the options.
A minimum of 50ng of RNA is required, however larger quantities are preferable to reduce the number of PCR cycles and therefore reduce duplicate sequencing reads. The protocol can be used with up to 1ug of total RNA, supplying at least 2µg of RNA would be ideal. Specialised protocols
Our standard protocol is designed to sequence RNA fragments with a length of approximately 200bp. If longer fragments are required please contact us to discuss the options.
Genomic DNA for whole genome sequencing NGS library preparation
Genomic DNA should be eluted in 10mM Tris-HCI (pH 7.5–8.5), low-EDTA TE buffer or nuclease free water. A minimum of 200ng of DNA is required; however, as with RNA-seq, larger quantities are preferable and supplying at least 1µg would be ideal. Protocols are available for smaller DNA quantities, please contact us to discuss the options. For all other NGS sample submission requirements please contact us.