- All specimens will be total RNA; prepared libraries; or genomic DNA from each Specimen to be analyzed.
- For DNA Sequencing (Whole-Genome Sequencing (DNA-Seq), Targeted Resequencing), EA requests using 1.5 ml non-stick tubes and a minimum 6.5 ug of genomic DNA at a concentration ≥ 125 ng/uL in a volume ≥ 25 uL in nuclease free water with an OD260/280 ratio of 1.8 – 2. If the Client is sending prepared libraries, EA requests using 1.5 ml non-stick tubes and all specimens should be ≥ 300 ng of 350 bp libraries at a concentration of ≥ 20 ng/ul at a volume of ≥ 15 ul. • DNA Sequencing and Targeted Resequencing: EA will process a PhiX control on all flow cells.
- For Exome Sequencing, EA requests 5 ug of total genomic DNA at a concentration of 500 ng/ul in Nuclease-free water (Agilent Exome Sure Select, 3 ug of gDNA minimum, Nimblegen SeqCap EZ Exome 1-5 ug of gDNA minimum). Note: all sequencing should be completed with paired end sequencing, 72 bp PE recommended. EA will process a PhiX Control specimen on all flow cells.
- For Small RNA Sequencing (miRNA-Seq), all specimens should be high quality RNA with RIN’s ≥ 7.0 and A260/A280 ratios ≥ 1.6. EA requests a minimum of 5 ug of total RNA at a concentration ≥ 125 ng/uL in a volume ≥ 25 uL in Nuclease-free water. Specimens should NOT undergo Lithium Chloride precipitation and should NOT contain tRNA or other nucleic acid co-precipitant. If Client provides prepared libraries, should be ≥ 300 ng of 350 bp libraries at a concentration of ≥ 20 ng/ul at a volume of ≥ 15 ul. EA will process a PhiX Control specimen on all flow cells.
- For RNA Sequencing (RNA-Seq), all specimens should be high quality RNA with RIN’s ≥ 7.0 and A260/A280 ratios ≥ 1.6. EA requests a minimum of 2.5 ug of total RNA at a concentration ≥ 125 ng/uL in a volume ≥ 25 uL in Nuclease-free water. Specimens should NOT undergo Lithium Chloride precipitation and should NOT contain tRNA or other nucleic acid co-precipitant. If Client provides prepared libraries, should be ≥ 300 ng of 350 bp libraries at a concentration of ≥ 20 ng/ul at a volume of ≥ 15 ul. A PhiX control is not required for this application. 72 bp PE sequencing (GAIIX) or 50 bp PE sequencing (HiSeq) is recommended for RNASeq if barcoding is required. EA will process a PhiX Control specimen on all flow cells.
- For ChIP-Seq, If EA is to prepare the library for the Client, EA requests using 1.5 ml non-stick tubes and a minimum of 150 ng of ChIP DNA at a concentration of 30 ng/ul in nuclease free water with an OD260/280 ratio of 1.8 – 2. If the Client is sending prepared libraries, EA requests using 1.5 ml non-stick tubes and should be ≥ 300 ng of 350 bp libraries at a concentration of ≥ 20 ng/ul at a volume of ≥ 15 ul. EA will process a PhiX Control specimen on all flow cells and recommends 36 bp SE sequencing. Note: Specimen precipitation must be completed before samples are shipped to EA. Alternatively, EA can have precipitation completed through one of our partner vendors. Please contact your Service Consultant for more information.
- For BiS-Seq (Bisulfate Conversion Sequencing), the Client is required to send prepared libraries. All specimens should be ≥ 150 ng of 350 bp libraries at a concentration of ≥ 10 ng/ul at a volume of ≥ 15 ul. EA requests using 1.5 ml non-stick tubes. EA will process a PhiX Control specimen on all flow cells.
- Our specimen requirements are designed to ensure that ample material is available for quantitation and quality assessment of the incoming DNA, with concentration values matched to the requirements of the assay. Adherence ensures that delays resulting from dilution or concentration of the sample are minimized. EA recognizes that these requirements may not be met in all situations. Your Service Consultant can work with you to accommodate special circumstances.
- After receiving the Specimen, EA will confirm amount and quality of Specimens using Agilent BioAnalyzer for prepared libraries or Agarose gel for genomics DNA, and PicoGreen analysis at a charge of $50 per specimen.