To improve the probability of having the highest quality of data we require a sample submission form from the user.  It is completely mandatory.

Sequencing Services:

• Option 1:  Full Service Sequencing
For Full Service Sequencing, the user provides the SGF with their quantified product (PCR or Miniprep products) and primer, each separately.  SGF will be charged of making all sequencing processing according to the users' quantified product and primer Ta.

Full Service Sequencing includes Post-PCR purification.  However, PCR products obtained with dyed PCR kits are not included and will not be considered for services.  So, please buy un-dyed PCR kits for your convenience.

We are currently using the PEG precipitation method for PCR clean-up; this clean-up is free of charge!  We are Not using Column Purifications or Exo-Sap, but these are available upon request and for an additional charge.

*Post-PCR purification will extend results' turn-around.


• Option 2: Ready to Sequence Service
  
  Ready to Sequence Services require that the user provides a Product-Primer mixture that will be ready to sequence and follows the following conditions:
• We recommend purifying your PCR template using Exonuclease I/Shrimp Alkaline Phosphatase (ExoSap) Digestion, Qiaquick column purifications and/or MiniPrep spin columns from Qiagen for plasmid DNA.  If using Qiagen columns, please elute with Autoclaved water.
• Please use strip-tubes or 96-well plates to send your samples.
• For PCR products, make a 7.0 uL mixture of:
  *15-100ng or 2ng per 100bp
  *1.0uL 3uM primer
  *bring volume up to 7.0uL with ddH₂O
  
  For plasmid DNA, make a 7.0 uL mixture of:
  *50-150ng of the insert+vector (miniprep product)
  *1.0 uL 3uM primer
  *bring up to 7.0uL with ddH₂O

For example: If your miniprep product has a concentration of 50ng/uL, your mixture should contain 3.0uL of your miniprep product (to have 150ng in your 7uL mixture), 1.0uL of your primer and 3.0uL of water.  You can now ship your mixture to the facility!

• Please contact us for BAC or Lambda DNA (or any other templates larger than 10kb).
  
• General notes:
  *We charge by multiples of 16 reactions.  If sending less reactions; make sure you have read the Terms of Agreement of the SGF.  Clients that send samples to SGF for processing will be considered to have read and fully agree with SGF Terms of Agreement.
  *Turn-around usually takes 24-72 hours depending on queue.  Longer turn-around will always be notified to the user.
  *We will gladly purify your product prior sequencing for an additional $20.00 per 96-well plate.
  *Positive controls are charged to your invoice.

Sample preparation for fragment analysis:

The most frequent source of problems prior most fragment analysis data collection is the DNA quality and concentration of the PCR reactions as it directly affects your genotyping run since a variety of contaminants such as RNA and proteins may be affecting your reaction.  This will result in poor band isolation and low yield bands.

For specific protocols to obtain any source of fragment analysis, please contact facility staff; they will be provided free of charge.  We are currently using a ROX based Size Standard for all fragment analysis protocols.

To solicit a quote for any fragment analysis/genotyping services, please e-mail us at sgf.upr@gmail.com!

Analysis Software Program availability:

Please visit our Bioinformatics Satellite laboratory (BSL) located at JGD-215.  This facility contains all the computing equipment necessary for the analysis of sequencing and genotype data.  Be sure to contact SGf personnel for Sequencher Key availability and any other information regarding data analysis.

• SGf is not responsible for data analysis or editing.
• We provide workshops for data analysis software program training as these are not included in our fees.
• Please contact SGF staff for workshop availability and for computer availability.

Ship samples to:

University of Puerto Rico, Río Piedras

Department of Biology

Julio García Díaz building-Room 214/216

Ave. Ponce de León

PO Box 23360

San Juan, PR 00931