Expression Labeling and Hybridization Service: Sample Submission Instructions
To facilitate processing of your order, please thoroughly review Sample Submission Instructions and prepare
samples accordingly. To ensure we receive what is needed for your order, please contact your NimbleGen
Account Manager with questions before preparing samples for submission.
Expression RNA Sample Preparation
1. High-quality total RNA (or Poly A RNA) is required to obtain optimally labeled transcripts
for assay hybridization.
2.
A suggested RNA isolation method successfully used with NimbleGen’s Expression Platform
is Qiagen’s Rneasy Kit Catalog #74104 or Trizol reagent.
3.
Samples should be concentrated to 1.0ug/ul in nuclease-free water via a standard sodium
acetate-ethanol precipitation.
4. Samples should be analyzed for quality and purity using Agilent’s Bioanalyser or
electrophoresis using a denaturing gel.
5. If DNA contamination is present, samples need to be treated with a standard DNAse
treatment prior to submission.
6. NGS Product Services Department will detection-label samples for hybridization.
7. NGS must receive 20ug of total RNA for Eukaryotic species submission. (If submitting poly
A RNA, NGS must receive 10ug per sample submission.) We approximate yielding enough
cRNA for 3 to 4 hybridizations from this amount.
8. NGS must receive 30ug of total RNA for Prokaryotic species submission. Due to the
Prokaryotic label method not including an amplification step, 30ug is requested per single
hybridization.
Quality Sample Requirements:
The customer before submission to NimbleGen Systems must perform this analysis:
1. RNA samples must have a concentration of 1ug/ul.
2. RNA samples must be submitted in nuclease free water.
3. Total RNA must give an OD260/280 ratio of at least 1.7.
4. RNA samples must be free of DNA contamination. (Please see following pages for
examples.)
5. RNA samples must show no signs of degradation. (Please see following pages for examples.)
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Sample Quality Examples:
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Sample Submission Process
Presubmission:
1.
Once your purchase order has been received, our internal account manager will contact you
to confirm service order details and for an estimated sample submission date. You will
receive a MANUFACTURING ORDER ID at that time. If you have multiple projects, each
project will receive its own manufacturing order ID.
2. Each unique sample should be submitted in its own tube. Please use skirted screw-capped,
1.5mL, micro-tubes for sample submission. Each tube needs to contain YOUR unique
identifier name for the sample. NGS Product Services Department will assign barcodes to
your samples upon receipt. Your unique sample identifier will also be tracked with the
sample along with NGS barcode number. Each tube must also contain your
MANUFACTURING ORDER ID.
3. Complete the Sample Submission Form (see Excel document) as specified.*
a. Each individual tube should have a line filled out on the Sample Submission Form
with that sample’s information.
b. The MANUFACTURING ORDER ID must be written on this form.
4. Complete the Sample Submission Checklist (see Expression Service Sample Checklist
document).*
a. The MANUFACTURING ORDER ID must be written on this document.
b. Please confirm checklist items have been completed.
5. *These two items must be filled out and submitted with samples for your order to proceed
without delays.
Post Submission:
1. Upon receipt by NimbleGen Systems, samples will be analyzed for RNA purity,
concentration, and quality (degradation) using the Agilent Bioanalyzer.
2. You will be contacted if there are discrepancies between NimbleGen Services sample
receiving QC and the data provided by you on the Sample Submission Form.
3. Samples that fail for suspected degradation or do not meet the requested amounts will not be
processed until you are contacted and an agreement is reached. In some cases, we may not be
able to process the samples due to insufficient amount.
4. Samples that require concentration changes may require additional fees to the order. *You
will be contacted before concentrations are corrected and fees are applied.
5. The option of resubmission is available for any samples that fail our incoming quality control.
Resubmission fees (to cover Quality control repeat on the resubmitted sample) will apply.
6. Any remaining Sample will be discarded 6 months after data delivery for your order.
If you would prefer to use samples again at a later date, please instruct account manager; storage fees
will apply.
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