| Service | Quantity | Harvard Internal | Harvard Affiliate/Academic | Commercial |
|---|---|---|---|---|
| 384-Well Standard Bulk Sample Order | 1+ | $4.12 | $4.74 | $5.36 |
| 96-Well Standard Bulk Sample Order | 1+ | $4.12 | $4.74 | $5.36 |
| Standard Individual Sample | 1+ | $4.12 | $4.74 | $5.36 |
Sanger DNA Sequencing
The BPF utilizes a local commercial contract research organization (CRO) to provide Sanger DNA Sequencing services to our core. The facility is located close by in Cambridge, Mass and provides sequencing under contract to a number of companies and institutes. The process for sequencing is quite simple. Samples are ordered through the BPF LIMS, tubes or plates are dropped off with requisitions in one of our freezer locations, samples are logged in by our core each morning and the courier picks up the samples from our core in late morning for processing at their laboratory on the same day. When ready, the results are quality checked by our core and data distributed within one business day of sample drop off through out LIMS.
Standard Sequencing Service
Users provide mixed template and primer in a tube or plate at the proper concentration. Samples have BigDye reaction chemistry, clean up and analysis on an ABI 3730xl instrument performed. Results are distributed to users in the form of .ab1 sequence chromatagram files. Sample tracking information in the form of date and time stamps can be viewed as the sample progresses from order to completion. Quality metrics for overall read length, Phred20 Read Length and average base quality score are provided. A sequence text and chromatogram viewer are also available directly through the website.Difficult Template Service
Identical to standard service but with alternate chemistry and cycling conditions to increase the likelihood of of sequencing success for templates with high GC, secondary structure, etc..The Biopolymers Facility provides DNA Sequencing services daily; Monday through Friday. The table below lists the deadline for sample drop off and the day the service will be performed.
| Drop Off Location | Drop Off Deadline | Services Timeline |
|---|---|---|
| New Research Building (NRB) | M-F before 9:00 AM | Data distributed by Noon next day (Tuesday to Saturday). |
| Goldenson Building | M-F before 8:00 AM | Data distributed by Noon next day (Tuesday to Saturday). |
| Building C-1 | M-F before 8:00 AM | Data distributed by Noon next day (Tuesday to Saturday). |
* When Harvard is closed the service deadline & execution will be the next business day.
Standard Sequencing and Difficult Template
- Microtube: 15µL or 30 µL *
- 96 well plate 15 µL total **
** For a 96-well plate submission, please plate the samples vertically by column (A01, B01, C01...H01 etc.)
Do not use TE Buffer, use instead 10mM 10 mM Tris pH 7-8 or Water.
In a microtube or plate prepare your samples with both template and primer according to the chart below:
| 15 ul | 30 ul | |
|---|---|---|
| PCR product | 50 ng | 100 ng |
| Double Strand DNA (plasmid) | 200 - 500 ng | 400 - 1000 ng |
| Single Stand DNA | 100 ng | 200 ng |
Primers should be pre-mixed with DNA templates: We recommend that you add 2 µl of 5 µM ( = 5 pmol/µl) primer per well or equivalent.
Arrange tubes by order number and snap closed
Line up samples tubes in the order in which they are listed on your requisition sheet. Seal tubes with caps.Labeling
Please label each tube only with the unique sample number listed next to the corresponding sample name on your requisition sheet. Each sample should have a unique number that follows the order your have arranged your tubes. Plates should be labeled with the requisition number.Drop off sample tubes in a BPF Freezer
See our list of Drop off locations.Standard Sequencing Service
Difficult Template Service
| Service | Quantity | Harvard Internal | Harvard Affiliate/Academic | Commercial |
|---|---|---|---|---|
| 384-Well Difficult Template Bulk Sample Order | 1+ | $14.00 | $16.10 | $18.20 |
| 96-Well Difficult Template Bulk Sample Order | 1+ | $14.00 | $16.10 | $18.20 |
| Difficult Template Individual Sample | 1+ | $14.00 | $16.10 | $18.20 |
We recommend viewing sequences with free software FinchTV or Chromas Lite if you are using a PC. If you have a Mac, we recommend using 4Peaks which is also freeware.