How to prepare samples

1) DNA or RNA SAMPLES for the preparation of libraries

  • Genomic DNA-seq protocol and targeted resequencing
  • Genomic DNA Mate Pair Protocol
  • ChIP-seq protocol
  • Protocol of RNA-seq (total RNA, mRNA)
  • SmallRNA-seq protocol
  • 16S rRNA sequencing protocol

2) SAMPLES of libraries already; ready for cluster generation and sequencing
3) Shipping of samples

Only the following samples are accepted for the performance of the NGS Bio-Fab service:

- DNA or RNA for library preparation, with quality requirements; and quantity as indicated on this page for each protocol (it is advisable to always wait for the indications of the laboratory, because requests may vary slightly depending on the application requested)
- Libraries already; ready for the cluster generation and amplification phase, and subsequent sequencing with the Illumina platform

1) DNA or RNA SAMPLES for library preparation

- Genomic DNA-seq protocol and targeted resequencing

  • Double-stranded DNA not degraded and not contaminated by RNA, proteins, salts and alcohols;
  • Quantity between 200 and 500ng total resuspended in dH2O (it is recommended to send the maximum quantity) at a concentration between 20 and 50 ng / ul;
  • 260/280 ratio equal to 1.8 and 260/230 ratio & gt; 2.

Recommendations: Spectrophotometric methods are not recommended for quantification, so & igrave; such as the Nanodrop, which cannot accurately discriminate between molecules of dsDNA, ssDNA, RNA and nucleotides present in the sample. It is necessary to evaluate the quality; and integrity of a DNA sample also through an electrophoretic run in agarose gel and attach to the sample sent an image of the run on the gel.

Notes: Bio-Fab guarantees the DNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

- Genomic DNA Mate Pair Protocol

  • Double-stranded DNA not degraded and not contaminated by RNA, proteins, salts and alcohols;
  • Minimum quantity to send: 10 & micro; g total resuspended in dH2O;
  • 260/280 ratio equal to 1.8 and 260/230 ratio & gt; 2.

Recommendations: Spectrophotometric methods are not recommended for quantification, so & igrave; such as the Nanodrop, which cannot accurately discriminate between molecules of dsDNA, ssDNA, RNA and nucleotides present in the sample. It is necessary to evaluate the quality; and integrity of the DNA running a small amount & agrave; of sample (200 ng) in a low percentage agarose gel (0.6%).

The DNA of excellent quality & agrave; it should correspond to a band generally greater than 50 kb with possible minimal presence of slight smearing at low molecular weight. If most of the DNA is less than 50 kb in size and the smearing is; clearly visible, the DNA & egrave; to be considered degraded. Attach a picture of the gel run to the submitted sample.

Notes: Bio-Fab guarantees the DNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

- ChIP-seq protocol

  • Immunoprecipitated double-stranded DNA, not degraded and not contaminated by RNA, proteins, salts and alcohols;
  • Minimum quantity to send: 10 ng immunoprecipitated DNA resuspended in dH2O (eluted in the smallest possible volume).

Notes: Bio-Fab guarantees the DNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST

- RNA-seq protocol (total RNA, mRNA)

  • purified total RNA not degraded and not contaminated by proteins, organic solvents and salts;
  • Quantity between 1 and 2 & micro; g total resuspended in nuclease-free dH2O (it is recommended to send the maximum quantity) at a minimum concentration of 100ng / ul;
  • 2100 Bioanalyzer RNA Integrity Number (RIN) = 8;
  • 28S rRNA: 18S rRNA = 2: 1
  • Ratio 260/280> 1.8

Recommendations: Sending quantities lower than those required can & ograve; lead to a reduction in yield in the preparation of the library, so & igrave; such as RNA degraded or contaminated with DNA molecules. So & egrave; extremely important to use high quality RNA; as a starting material. It is recommended to include a step in DNase during the RNA isolation protocol. DNA contaminants can however be removed during mRNA purification.

It is necessary to evaluate the integrity; of RNA by running it in an agarose gel with 1% formaldehyde and staining with ethidium bromide. High quality RNA shows a 4.5 kb band corresponding to the 28S rRNA. This band should have an intensity; twice as much signal as shown by the 1.9 kb 18S rRNA band. Attach a picture of the gel run to the submitted sample.
If you decide to start with mRNA rather than total RNA, it is; It is necessary to use the purified mRNA fraction from 0,1 & ndash; 4 & micro; g of total RNA.

Notes: Bio-Fab guarantees the RNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

- SmallRNA-seq protocol

  • purified total RNA not degraded and not contaminated by proteins, organic solvents and salts;
  • Minimum quantity to send: 1 & micro; g total resuspended in nuclease-free dH2O;
  • 2100 Bioanalyzer RNA Integrity Number (RIN) = 8;
  • 28S rRNA: 18S rRNA = 2: 1
  • Ratio 260/280> 1.8

Recommendations: Sending quantities lower than those required can & ograve; lead to a reduction in yield in the preparation of the library, so & igrave; such as RNA degraded or contaminated with DNA molecules. So & egrave; extremely important to use high quality RNA; as a starting material. It is recommended to include a step in DNase during the RNA isolation protocol. DNA contaminants can however be removed during mRNA purification.

It is necessary to evaluate the integrity; of RNA by running it in an agarose gel with 1% formaldehyde and staining with ethidium bromide. High quality RNA shows a 4.5 kb band corresponding to the 28S rRNA. This band should have an intensity; twice as much signal as shown by the 1.9 kb 18S rRNA band. Attach an image of the gel run to the submitted sample.
If you decide to start with small RNA rather than total RNA, it is; It is necessary to use the smallRNA fraction purified from 1 -10 μg of total RNA.

Notes: Bio-Fab guarantees the RNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

- 16S rRNA sequencing protocol

  • DNA not degraded and not contaminated by RNA, proteins, salts and alcohols;
  • Quantity of about 500ng total resuspended in dH2O (it is recommended to send the maximum quantity) at a minimum concentration of 50ng / ul;
  • 260/280 ratio equal to 1.8 and 260/230 ratio & gt; 2.

Recommendations: Spectrophotometric methods are not recommended for quantification, so & igrave; such as the Nanodrop, which cannot accurately discriminate between molecules of dsDNA, ssDNA, RNA and nucleotides present in the sample. It is necessary to evaluate the quality; and integrity of a DNA sample also through an electrophoretic run in agarose gel and attach to the sample sent an image of the run on the gel.

Notes: Bio-Fab guarantees the DNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

2) Libraries already ready for cluster generation and sequencing

  • Quantity: 10 & micro; l of a 10nM concentrated dH2O solution;
  • Always check compatibility; of the indexes used

Recommendations: Sending quantities lower than those required can & ograve; lead to a reduction in cluster formation. So & egrave; It is extremely important to determine the concentrations of the samples that can be obtained by reading the QuBit, Bioanalyzer and qPCR (as described in the ILLUMINA qPCR Quantification Protocol Guide).

Notes: Bio-Fab guarantees the DNA quantification service for this protocol. For the cost of the service consult the NGS PRICE LIST.

3) Shipping of samples

Samples must be shipped in 0.2 ml DNase / RNase free tubes, sealed, sequentially numbered and individually named with the names of the samples they contain. It is recommended to ship frozen (DNA) or dry ice (RNA) samples by express courier, with guaranteed delivery within 24 hours.

Shipping of samples or library is; always subsequent to the acceptance of the service access form by Bio-Fab, and subsequent to the acceptance of the conditions of sale by the customer.

To send the samples and for any further information send an email to This email address is being protected from spambots. You need JavaScript enabled to view it.

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