Sample Preparation

Samples should be delivered to the UNMC Genomics Core Facility room 1010 DRCII.  Please see below for instructions regarding how to prepare the samples prior to bringing them to the facility.

Sample Preparation

For all samples, use 0.2 mL PCR 8 tube-strips.  For orders of more than 48 samples, you should use a 0.2 mL PCR plate sealed with 8-cap strips; start at well A1 and go down the columns vertically.

Jump to specific instructions for:

• Premixed Samples
• Predefined Samples
• Custom Samples
• Unpurified PCR Products

Premixed samples

For Premixed samples, formulate each sample in two parts:

Add the required amount of "DNA total mass" to the sample tube and then add enough water to make a total volume of 100 μL.

Make a primer stock of 5 μM primer.  Add 5 μL of the appropriate primer stock (5 μM) of your primer to the sample tube to make a total sample volume of 15 μL.

if you have chosen to use a primer provided by Genewiz, submit the 10 μL DNA sample.  Genewiz will add the primer.

Premix Sample Table:

Predefined Samples

For Predefined samples, you provide your DNA samples and primers in separate tubes.  Genewiz will calculate and dilute your sample and add the appropriate amounts of DNA and primer to each of your sequencing reactions.  You must provide an accurate DNA concentration for each of your samples.  Each of your primers should be in a separate tube at a concentration of 5uM; you will need to include enough primer for 5uL per reaction.  For Predefined samples, you need to provide the DNA at a mass amount equal to or greater than the concentrations and mass amounts shown in the table for Predefined samples. 

 Predefined Sample Table:

Custom Samples

For Custom samples, Genewiz will measure the concentration of your DNA.  Genewiz will also dilute your primer based on the primer concentration information you provide.  They will then add the appropriate amount of DNA and primer to each sequencing reaction.  For Custom samples, you need to provide the DNA at an estimated concentration and mass amount equal to or greater than the concentrations and mass amounts shown in the table for Predefined samples.

Unpurified PCR Products 

Unpurified PCR products can be sent to Genewiz for sequencing for an extra charge.  Genewiz will purify your DNA using an Exo-SAP treatment.  Please provide more than the required amount of PCR DNA according to the tables above for Predefined sequencing samples. Also, provide 5uL per sequencing reaction of each primer at 5uM in separate tubes. PCR products should show only one band on a gel for this service to be appropriate.  You need to provide an estimated concentration for the PCR DNA in your sample.  Also, provide a picture of a gel of the sample with labels to indicate lanes for each sample, and provide information on volume of sample loaded, volume of ladder and information on the ladder, to allow Genewiz to check concentrations of samples using the gel.  Concentrations of unpurified PCR DNA products cannot be measured on a Nanodrop or other spectrophotometer, including after Exo-SAP treatment.  Please send your gel picture and info to us along with your order form, by email at SangerSequencing@unmc.edu