Protocols

Sample requirements

RNA

Quantity 5- 6 µg resuspended in nuclease-free dH2O (we recommend sending the maximum amount).
Purity: ratio 260/280 of 1.8 or higher; 1.8 < 260/230 < 2.1
Integrity: Bioanalyzer RNA Integrity Number (RIN) >= 8*

*If you can't perform Bioanalyzer analysis it is necessary to evaluate the integrity of the RNA in running agarose gel with 1% formaldehyde and staining with ethidium bromide. Please attach a picture of the run of the gel to the sample sent.

NOTE: In case of genomic DNA contamination the RNA sample must be treated with DNAse and purified before sending.


gDNA

Quantity 6-10 µg resuspended in nuclease-free dH2O (we recommend sending the maximum amount)
Purity: ratio 260/280 of 1.8 or higher; 1.8 < 260/230 < 2.1
Integrity: It 's necessary to evaluate the quality and integrity of a DNA sample through a run in 0.7% agarose gel and please attach a picture of the run of the gel to the sample sent.**

**A DNA of good quality should correspond to a band generally greater than 50 kb with minimum possible presence of slight smearing of low molecular weight. If most of the DNA shows a size less than 50 kb and the smearing is clearly visible, the DNA is considered to be degraded.


gDNA for Mate Pair libraries

Quantity for 2-5 Kb libraries: 25 µg resuspended in nuclease-free dH2O
Quantity for 5-10 Kb libraries: 45 µg resuspended in nuclease-free dH2O
Purity: ratio 260/280 of 1.8 or higher; 1.8 < 260/230 < 2.1
Integrity: It 's necessary to evaluate the quality and integrity of a DNA sample through a run in 0.7% agarose gel and please attach a picture of the run of the gel to the sample sent.**

**A DNA of good quality should correspond to a band generally greater than 50 kb with minimum possible presence of slight smearing of low molecular weight. If most of the DNA shows a size less than 50 kb and the smearing is clearly visible, the DNA is considered to be degraded.

NOTES: Sending sample amount lower than those required and/or low quality may result in a reduction of library quantity/quality and compromise the clusters formation. So it is extremely important to determine the concentrations of samples obtainable by Fluorometer/ Spectrophotometer and Bioanalyzer.

Sending samples

Shipment of samples or the library is always after acceptance of the submission form to the Functional Genomics Lab.

Samples should be shipped as acqueous solution in dry ice in tubes of 1.5 ml DNase / RNase free, sealed and named (legibly) individually with the names of the samples and of the project manager.

In case the shipment on dry ice is not possible samples may be shipped precipitated in ethanol:

  • Prepare DNA/RNA water solution with total volume of not less than 30 μl.
  • Add 0.1 volumes of 3 M sodium acetate and 2-3 volumes of ice cold 100% ethanol and mix (please use DNAse free reagents).

Please ensure that the total volume of sample solution is not less than 120 μl. To prevent DNA/RNA degradation in shipment, please do not centrifuge samples after ethanol precipitation prior to sending and send the sample in screw-capped tubes to prevent alcohol evaporation (it's important!).

Please send your samples to the following address:
Department of Biotechnologies, University of Verona
Strada le Grazie, 15 - Cà Vignal, 1 - 37134 (VR) - Italy
Phone: +39 045 802 7058
Fax: +39 045 802 7929


Last modified: Wednesday October 08, 2014

Functional Genomics Lab

Department of Biotechnologies, University of Verona
Strada le Grazie, 15 - Cà Vignal, 1 - 37134 (VR) - Italy
Phone: +39 045 802 7058
Fax: +39 045 802 7929