-Requires 5-10 micrograms or more at minimum concentration of 100ng/ul
-Total RNA must be of high integrity (RIN > 8.0)
-Requires 500 nanograms to 1 microgram or more at minimum concentration of 50ng/ul
-Method can be compatible with degraded RNA, so long as each sample within the experiment is similarly degraded
-Requires 500 nanograms to 1 microgram or more at minimum concentration of 50ng/ul
-Method can be compatible with degraded RNA, so long as each sample within the experiment is similarly degraded
-Requires 500 nanograms to 1 microgram or more at minimum concentration of 56ng/ul
-Method can be compatible with degraded RNA, so long as each sample within the experiment is similarly degraded
-Requires 10 nanograms or more at minimum concentration of 1.1ng/ul
-Total RNA must be of high integrity (RIN > 8.0)
– Requires 10 nanograms or more at minimum concentration of 1.2ng/ul, for FFPE recommended input is 50 nanograms at minimum concentration 6ng/ul
-Method is compatible with degraded RNA, particularly FFPE-derived samples, so long as each sample within the experiment is similarly degraded
-If total RNAs are of good integrity, we recommend Clontech for low input RNA-seq
-Requires 600 nanograms to 1 micrograms or more
-Requires 100 to 500 nanograms or more, coverage dependent
-Requires 5 to 100 nanograms or more
-Requires 100-200 nanograms or more
-Requires 200 nanograms or more
Please contact the GTAC@MGI prior to sample preparation regarding specific plate types, lysis buffer, RNase inhibitor, and volumes required for these services.
This method will provide sequence data that is strongly 3’ biased due to the location and detection of the barcodes added during cDNA generation. Because of this we are only able to provide gene level data, not transcript level.
Please Note: FACSseq libraries are not compatible with our standard dual-indexed NovaSeq S4 (2×150) sequencing workflow. Sequencing instrumentation and run type will be determined based on the size of the project and desired read-depth. The GTAC@MGI will confirm run type when samples are received. Targeting 1 to 2 million reads per cell is recommended.
Assuming adequate sample quality (RNA is highly intact) the SMARTseq method generates sequence data across full transcripts, and typically requires greater sequencing depth compared to FACSseq to detect similar numbers of expressed genes per cell. These libraries are dual-indexed and sequenced in our standard Illumina NovaSeq S4 (2×150) workflow. Targeting 5 to 10 million read-pairs per cell is recommended.
FACSseq and SMARTseq2 library preps are charged by the (96-well) plate. If partial plate is processed, there is no discount for unprocessed wells.
Sample submission requirements: Contact PacBio Production Manager Chris Markovic (cmarkovi@wustl.edu)
Sample submission requirements: Contact Oxford Nanopore Production Manager Chris Markovic (cmarkovi@wustl.edu)
Sample submission requirements: Contact Bionano Production Manager Chris Markovic (cmarkovi@wustl.edu)
Sample submission requirements: Contact Lab Research Manager Chris Sawyer (csawyer@wustl.edu)
GTAC@MGI requires all samples be submitted with the correct submission form and template matrix. Samples must be randomized across the 96 sample matrix. Samples received without submission forms, or forms missing required information will not be processed until complete information is provided.
Samples should be provided in well labeled 1.7ml tubes, with the unique label matching the sample name on the submission form. As well, please include PI and date. Please provide 150 µl aliquots for each sample.
NOTE: Improperly prepared samples may impact analyte measurements. In addition, light scattering can be indicative of particulates of sufficient size to clog filters during a SOMAscan Assay run, which could lead to a failed sample run.
Prior to processing samples through the SomaScan assay, GTAC microarray will evaluate all submitted protein. Please review SomaScan Sample Handling. Initial QC specifications will require an email confirmation if you wish to proceed with processing. If a sample should fail due to a technical error, investigators will only be charged for the samples that were completed.
Agilent SOMAScan slides will be processed through the Agilent scanner and image data will be extracted using Agilent Feature Extraction software. Extracted initial raw signal intensity data will be uploaded onto the SOMAScan server for data QC and standardization. The final data set will include SOMAscan generated .adat file, a reformatted text file with standardized relative fluorescent unit (RFU) for ~7K proteins, SOMAscan Quality Statements (SQS) and basic differential expression.