The UNLV Genomics Core Facility offers Sanger DNA sequencing services. Sanger sequencing is run using an either an ABI 3130 or ABI 3500 Dx genetic analyzer and ABI BigDye v3.1 Terminator sequencing chemistry. The typical contiguous read length is > 900 bp, with QV's >20 for high quality, clean DNA templates. The typical turn-around time is two to four business days, although this may change depending on our sample workload.
Sample Submission Information
To submit samples for sequencing, please provide the appropriate amount of DNA template (see the chart below) and primer in 0.2 ml flat-cap PCR tubes (no strip tubes please) in a final volume of 10 ul of nuclease-free water. Please do not submit templates in TE buffer for the EDTA will chelate the metal ions (Mg++) required for the cycle sequencing reaction. If you are adding your own primer, please make the final concentration between 3.2 and 10 uM of primer. The required template input amounts are:
|PCR product: 100 to 200 bp
||1 to 3 ng
|PCR product: 200 to 500 bp
||3 to 10 ng
|PCR product: 500 to 1000 bp
||5 to 20 ng
|PCR product: 1000 bp to 2000 bp
||10 to 40 ng
|PCR product: >2000 bp
||40 to 100 ng
|Bisulfite converted genomic DNA-PCR product
||3 to 10 ng
||50 to 100 ng
||200 to 500 ng
||0.5 to 1.0 ug
|Bacterial genomic DNA
||2 to 3 ug
Submitted samples must be clearly labeled in 0.2 ml flat-cap PCR tubes and each sample must have a unique identifier. The requestor must complete a DNA Sequencing Sample Submission Form and send it by email to Casey Hall-Wheeler at: firstname.lastname@example.org before bringing sequencing samples to the Genomics Core.
Measures must be taken by the investigator to provide the cleanest DNA template possible for the overall success of the sequencing reaction and data is highly dependent on the quality and quantity of the DNA sample. A pGEM 3Zf(+) control is run with every sample set to ensure instrument and technical performance. Unincorporated dye terminators are removed from the extension products with using the BigDye XTerminator Purification Kit. We can use specialized cycle sequencing and genetic analyzer run protocols for difficult to sequence templates including short PCR products, GC-rich templates, or sequencing primers with non-standard annealing temperatures (inquire for details).
We run sequencing Monday through Thursday by scheduled appointment only. Our turn around time is between two to four days from the time you submit your samples until you have data in hand, although this timing is dependent on our current workload. To schedule an appointment for DNA sequencing services, please call Casey Hall-Wheeler at the UNLV Genomics Core at 702-895-1057.
Universal Sequencing Primers Offered by the UNLV Genomics Core
The UNLV Genomics Core offers the following sequencing primers: T7, SP6, pBABE3', pBABE5', pGEX3', pGEX5', CMV forward and M13 reverse. The sequences for these primers are:
||5'- AATACGACTCACTATAG -3'
||5'- GATTTAGGTGACACTATAG -3'
|(-21) M13 Forward
Cost for DNA Sequencing
The fee for DNA sequencing for samples that are ready to run and do not need to be manipulated by us is $6.25 per sequencing reaction. The fee for DNA sequencing for samples that need to be manipulated by us (addition of primer, verify template concentration on the Nanodrop, transfer submitted samples that are not in 0.2 ml flat-cap PCR tubes into 0.2 ml flat-cap PCR tubes) is $7.50 per sequencing reaction. For a complete list of our DNA sequencing pricing, please see our Fees page.
More information regarding our DNA sequencing services can be found at the following links: