Exome Sequencing

 

Exome sequencing using Ion Proton sequencer (Life Technologies)

Method of sample preparation

DNA extraction using commercial kits is recommended. Please indicate the method used for the extraction/purification and the buffer for final elution. Low salt buffers or water are preferred.

Quantity and quality

500 ng of high-quality gDNA must be supplied in a volume between 15-50 μl and  should be quantified with Qubit DNA HS Assay Kit (otherwise, please indicate the alternative method used). With the spectrophotometer, the A260/A280 ratio should be between 1.8 and 2, and the A260/A230 one between 2 and 2.2.  Please indicate the method used for the extraction/purification and the buffer for final elution. Low salt buffers or water are preferred. A generic PCR amplification to assess the template quality is recommend.

Service Description

Library preparation starting from gDNA samples with the Ion AmpliSeqTM Exome Kit, sequencing with  Ion Proton Sequencer and bioinformatic analysis. The guaranteed coverage is 80X.

The bioinformatic Torrent SuiteTM is used to generate Alignment BAM files, Coverage Analysis statistics and Variant Analysis VCF files. By default the Variant calling is performed using the “Germ Line-High Stringency” algorithm. Write us if your data require a different algorithm.

Results are saved on an hard disk and returned to customer by express delivery.

Sample and data storage

After data delivery, the sample and the data are stored 30 days, unless CRIBI and the customer have defined a different agreement.

Sample shipment

We recommend to send the sample in 1.5 ml tubes (preferably low binding). Please write clearly the sample CODES  associated by our booking system, the concentration and the volume on the tubes. SAMPLES WITHOUT CODES ARE NOT ACCEPTED.

DNA can be sent in styrofoam containers with ice packs or dry ice.

Send the sample to the address below:

 

CRIBI Proton Service

Att.ne Dr. ssa Michela D’Angelo

Università di Padova

Complesso Biologico Vallisneri

via Ugo Bassi, 58/B

35121 Padova (Italy)

ngs@cribi.unipd.it