Genetic Sequencing

MiSeq Cluster

This page and the links listed have been designed to provide students and researchers who are interested in genetic sequencing with information on the sequencing available at the University, and how it works. In addition we aim to provide you with the information you need to select the correct technology for your project and how to submit your DNA samples.

Available Sequencing Techniques

Quick Facts

  • NGS funded by the NSF
  • The cutting edge next level of genetic sequencing to generate meta-data
  • For sequencing of prepared NGS libraries
  • Metagenomic reports generated with 16S analysis

The Miseq from illumina is used in cutting edge research and increasingly in healthcare for applications such as personalized medicine, IVF, cancer research. We can provide specialist advice if you have a project in mind and are not sure how to generate the data. In order to provide an estimate for cost per sample or per project please contact biology+tech@montclair.edu. For 16S metagenomic analysis we will provide a pdf doc per sample as generated by the onboard program that compares the sequencing to the greengenes database.

View a sample report

Quick Facts

  • The gold standard for Genetic Sequencing
  • Keywords: PCR product, plasmid
  • Max length generated 750-850bp

Capillary electrophoresis (Sanger) Sequencing is still widely used and accepted as the gold standard of genetic sequencing due to its very high accuracy and has been used in research for the past 20 years. The process involves three stages once the sample is ready to be sequenced:

  1. A cycle sequencing reaction where the fluorescently tagged nucleotides are incorporated into any DNA present by using a specific primer
  2. The whole reaction must be cleaned up to remove all impurities
  3. The sample is loaded onto the 3130 genetic analyzer where capillary electrophoresis produces an electropherogram which contains your genetic sequence.

Read through the Applied Biosystems Sequencing Handbook for information on our sequencer.

Sequencing Samples Submission

General Requirements

Submit sequencing samples according to the instructions below:

  • Email biology+tech@montclair.edu a gel pic with the indicated bands and a positive and negative control if possible
  • Provide a labeled tube for the PCR product or plasmid (label plasmid tube with conc. in ng/µL) and a labeled tube for the forward and/or reverse primer (do not combine primers). Primer concentration should be 10µM
  • If you are setting up your own sequencing (ONLY for PCR) pre-mix. Use 1.5µL PCR, 1.5µL single Primer (At concentration of 10µM) and 7µL water for a total of 10µL
  • Clearly indicate a unique name for each sample and clear instruction if various primers are to be used
  • Use the freezer in REID 310B marked ‘Sequencing Samples’ to transfer your samples and email biology+tech@montclair.edu when you leave samples

Quick Facts

  • Mainly for population variance or progeny verification
  • Keywords: RFLP, microsatellites, STR

Fragment analysis is an additional way to analyze genetic material. It has a wide range of applications such as:

  • Studying the variation of individuals in a  population using amplified fragment length polymorphisms (AFLP).
  • Comparing known genetic profiles to identify individuals by examining microsatellites amplified by fluorescently tagged primer. Used by law enforcement to identify suspects also used in paternity testing (Heredity).
  • Analysis of single nucleotide polymorphisms (SNPs) using SNPlex or SNaPshot. Processes generally used to identify single base pair mutations at a specific site, like cancer or genetic condition screenings.

Pricing can be drastically different depending on the needs of the associated project, please contact biology+tech@montclair.edu for further information.