Sample Requirements
To ensure the best sequencing results, please follow the guidelines below when submitting your Premium PCR samples:

• Sample Type: Linear, double-stranded DNA only (single-stranded DNA may result in unpredictable results).
• DNA Purity: DNA should be purified using a column-based method (e.g., Zymo Clean & Concentrator) or magnetic beads (e.g., Ampure XP). Unpurified DNA will not be accepted.
• Volume: A minimum of 10 μL per sample is required, submitted in 200 μL strip tubes.
• Buffer: Use elution buffer (10 mM Tris, pH 8.5) or nuclease-free water. Avoid using buffers containing DMSO.
• Concentration: For optimal sequencing results, send your samples at the following concentrations based on the size of your DNA insert:
• 100 bp: 1 ng/μL
• 200 bp: 2 ng/μL
• 500 bp: 5 ng/μL
• 1000 bp: 10 ng/μL
• 2000 bp: 20 ng/μL
• 5000 bp: 50 ng/μL
• 10000 bp: 100 ng/μL
• 15000 bp: 150 ng/μL
• 20000 bp: 200 ng/μL
• 25000 bp: 250 ng/μL

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Verify Concentration

Please provide your Premium PCR samples at the recommended concentration and minimum volume listed in the table above. For optimal sequencing performance, we highly recommend measuring DNA concentration using a Qubit or another fluorometric method (such as a plate reader), as these provide the most reliable results.

If you choose to use spectrophotometric methods like Nanodrop, keep in mind that these tend to be less precise for DNA quantification. Samples submitted at concentrations that are too high or too low can compromise library preparation and sequencing, potentially leading to unsuccessful runs. In fact, incorrect sample concentration—often caused by reliance on Nanodrop readings—is the most frequent reason for sequencing failure. Careful quantification and proper normalization are essential for consistent, high-quality results.

Verify Quality

This service is designed specifically for linear, double-stranded DNA. We suggest confirming the size of your full-length DNA molecules using gel electrophoresis before submission.

Verity Purity

We recommend that DNA samples have a 260/280 ratio above 1.8 and a 260/230 ratio between 2.0 and 2.2. Purity can be assessed using Nanodrop or other spectrophotometric methods.

To ensure optimal sequencing performance, samples should be free of:

• RNA (treat with RNase during extraction if needed)
• Denaturing agents (such as guanidinium salts or phenol) and detergents (like SDS or Triton-X100)
• Residual biological contaminants from the source organism (e.g., heme, humic acids, polyphenols)
• Insoluble particles, visible coloration, or cloudiness