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Sanger Sequencing Sample Requirements

The RTSF Genomics Core can provide short turnaround time on small and high-throughput projects utilizing the 96 capillary electrophoretic ABI 3730xl platform. The expected sequence length of PGEM plasmid controls typically ranges from 800 to 900 bp.

photo of well plates

Service Options

  • Custom (Individual Tubes, <= 16 samples)
  • High-Throughput (8-strip(s) for 8-16 samples, 96-well plate(s) for >16 samples)

Types of Reactions

  • Standard Reaction
  • 1st Tier Reaction - for high GC content and plasmids > 10kb
  • Genomic Reaction - for genomic/chromosomal DNA
  • GTP Reaction - for siRNA hairpins and secondary structure

Sample Requirements

  • The volumes requested below are sufficient for two reactions.  If less volume is provided, samples cannot be resequenced if the initial reaction fails.
  • For PCR products, it is very important to purify your sample to remove any remaining primers from the reaction, unincorporated nucleotides, enzyme, buffer, and salts.  Make sure any residual ethanol is removed as well.  Options for purification include: bead or column-based purification, enzymatic cleanup, gel extraction, and ethanol precipitation. 
  • Sample concentrations should be measured by fluorometric method (such as Qubit or PicoGreen).  UV-based measurements (such as NanoDrop) underestimate sample concentrations and should not be used.
  • Samples should be submitted in sterile water.
  • Add only one primer to each sample.  If you would like to sequence with the forward and reverse primer, two reactions are necessary.

For samples with client-added primer:

DNA Type

DNA Total Mass (ng)

Primer to add (picomoles) Total Volume (ul)
Plasmids
single-stranded DNA 200 30 12
double-stranded DNA (up to 10 kb) 1000 30 12
Purified PCR Product
<100 - 200 bp 2 - 6 30 12
200 - 500 bp 6 - 15 30 12
500 - 1000 bp 10 - 40 30 12
1000 - 2000 bp 20 - 80 30 12
>2000 bp 80 - 200 30 12

 

For samples requesting the addition of a Genomics Core standard primer:

DNA Type

DNA Total Mass (ng)

Primer to add (picomoles) Total Volume (ul)
Plasmids
single-stranded DNA 200 0 8
double-stranded DNA (up to 10 kb) 1000 0 8
Purified PCR Product
<100 - 200 bp 2 - 6 0 8
200 - 500 bp 6 - 15 0 8
500 - 1000 bp 10 - 40 0 8
1000 - 2000 bp 20 - 80 0 8
>2000 bp 80 - 200 0 8

Sample Submission Overview

  • We accept purified plasmids and purified PCR products.  All samples must be submitted with a completed submission form.  Submission forms can be downloaded from our Forms Webpage.
  • Sample requirements are provided in the tables above.
  • Six standard primers are offered by the Genomics Core at no additional charge.  If you will not use one of the standard primers, the customer must add their own primer to each sample.
  • For partial plate submissions, please arrange samples by column (i.e., A1-H1, A2-H2, etc.), NOT by row. A $20 re-racking fee will be charged for any samples not arranged in columns.
  • Sequences can be accessed and downloaded from the Genomics Depot. If your data is not available after 48 hours please contact us at gtsf@msu.edu.  Data is only available for 6 months.  See our Data Retention Policy webpage for more information.

Sample Drop-off

The Genomics Core is accepting samples for all projects, but because of restricted access to the Plant Biology Laboratories building, researchers who do not otherwise have access to the building must drop-off samples at the Genomics Core refrigerator at the loading dock of the Plant Biology Laboratories building. Researchers should send an email to gtsf@msu.edu to indicate that their samples have been deposited in the loading dock refrigerator. 

Researchers who otherwise have access to the Plant Biology Laboratories Building may continue to drop off samples in the hallway refrigerator outside of the Genomics Core (room S-18).

Due to the pandemic, the drop box located on the 5th floor of BPS is closed.

Plant Biology building map showing the loading dock

 



Shipping samples

Off-campus researchers should be aware that “overnight deliveries” from both FedEx and UPS are often taking two days.  These overnight packages are being delayed in warehouses outside of Michigan.  Once packages arrive in East Lansing, they are being delivered correctly, and we are having no problems with packages that make it to our building.  Because of the probability of delay, samples should be sent with a gross excess of dry ice.  We recommend five pounds or more.  Additionally, plastic film seals for 96-well plates should be avoided as melted samples readily leak into adjacent wells.  To avoid leakage, we recommend an excess of dry ice and using strip caps that were designed for the 96-well plate or sealing foil.

Address shipments to:

Colleen Curry
612 Wilson Road, Rm S-18 D
East Lansing, MI 48824