| Service | Quantity | Harvard | Non-Harvard/Academic | Commercial |
|---|---|---|---|---|
| Standard Individual Sample (1 to 24) | 1-24 | $8.50 | $9.78 | $11.06 |
| Standard Individual Sample (25 to 48) | 25-48 | $7.50 | $8.63 | $9.76 |
| Standard Individual Sample (49 or more) | 49+ | $5.75 | $6.62 | $7.48 |
| 96-Well (1 Plate) | 1 | $4.50 | $5.18 | $5.86 |
| 384-Well (1 Plate or more) | 1+ | $2.00 | $2.30 | $2.60 |
| 96-Well (2 Plates) | 2 | $4.00 | $4.60 | $5.20 |
| 96-Well (3 Plates) | 3 | $3.50 | $4.03 | $4.56 |
| 96-Well (4 Plates or more) | 4+ | $2.00 | $2.30 | $2.60 |
Sequencing
DNA sequencing is the process of determining the order of nucleotide bases in a particular DNA molecule. With our automated DNA Sanger sequencing pipeline, we can process both purified as well as unpurified samples with a typical turnaround time between 24-48 hours for most samples.
Standard Service
The standard DNA sequencing service at the Biopolymers Facility begins with a cycle sequencing reaction using Dye Labeled Terminators to generate extension products from client template DNA. The reaction is then purified using a magnetic SPRI chemistry before being processed on a 3730XL DNA Analyzer from Applied Biosystems. All sequencing data is checked using our quality control software.PCR Clean
We offer a PCR purification service using magnetic SPRI chemistry to remove PCR primers before sequencing. Unpurified PCR products may be submitted in half-skirted 96-well PCR plates. A PCR reaction with volumes between 20µL to 50µL can be submitted for PCR cleanup. Sequencing primers must be provided by the client in a standard 1.5µl micro-tube in µM concentration with a total volume of at least 500µL. The client may provide as many as two primers per plate being purified. The first primer will be used in a sequencing reaction included in the cost of the PCRClean service. Please note that if a second primer is submitted, it will need a separate requisition for "standard bulk sample order".SEQClean & Analysis
If you have already performed cycle sequencing reaction using dye labeled terminators (BigDye v3.1) and would like us to purify and run the samples, you can select our "SEQClean & Analysis" service. The reaction is purified using a magnetic SPRI chemistry before being processed on our 3730XL DNA Analyzer from Applied Biosystems. All sequencing data is checked using our quality control software.3730xL Analysis Only
If you have already purified the dye labeled fragments and would like us to sequence the samples, you can select our 3730xL Analysis Only service where we will take your plate and run it on our 3730XL DNA Analyzer from Applied Biosystems. All sequencing data is checked using our quality control software.The BPF is currently equipped with:
- Applied Biosystems 3730XL DNA Analyzer
- Hamilton Robotics Microlab StarPlus liquid handler
- Velocity 11 PlateLoc
Sample Requirements
PCR Clean- 500ul of 1uM primer (separate tube)
- 20-50ul PCR product
- 0.5 ml microtube: 15ul or 30ul
- PE style half skirt plate: 15ul total*
- PE style half skirt 96-well plate: 10ul total labeled fragments
- PE style half skirt 96-well plate: 10ul purified labeled fragments
| 15 ul | 30 ul | |
|---|---|---|
| PCR product | 12.5 ng/100 base pairs | 25 ng/100 base pairs |
| Double Strand DNA (plasmid) | 500 ng | 1000 ng |
| Single Stand DNA | 100 ng | 200 ng |
| Note: Contact us for details on BAC sequencing | ||
| Primer | 4 pmol | 8 pmol |
* Custom 0.5 ml DNA Sequencing tubes may be purchased through our Reagents & Supplies program, under Sequencing Supplies.
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.Label tube with sample number
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. Sample names should not be written on tubes.We highly recommend using the labels we have in stock for tube labeling.
Drop off sample tubes in a BPF Freezer
See our list of Drop off locations.Basic Sequencing
All samples for our basic sequencing service must be submitted in custom 0.5 ml microtubes. You may purchase our custom 0.5 ml tubes through our Reagents & Supplies program, under Sequencing Supplies. The minimum volume is 15ul including template and primer. For concentration information and step-by-step submission instructions please follow the links above for your preferred submission size.You may also submit samples for basic sequencing in 30ul volumes, which will automatically qualify your samples for free reruns in the event of a failed reaction. Again please see the details above for concentration information.
Pre-Sequencing and Partial Services
Samples submitted for Whole Plate Orders receive our discounted ordering price and are eligible for pre-sequencing or partial service options. Follow the link above for whole plate instructions regarding purification of PCR products and plasmids as well as BigDye cleanup and 3730xL Analysis. Partial plates are acceptable as whole plate orders but will be billed as a full unit.Standard Primers
The Biopolymers Facility requires all users to provide their own primer for each DNA sequencing sample. If you need a standard primer such as T7 you can order it through our oligonucleotide synthesis service and the primer will be available here for pick up. For primer design hints and primer sequences see follow our information link above.
Our basic sequencing service includes dideoxynucleotide terminator PCR and reaction cleanup, along with capillary electrophoresis and signal
analysis. Discounts are available for bulk orders and will be billed at the discount price x 96. Bulk sample sets will be assigned one full 96-well
PCR plate for reactions and analysis. There is no need to fill a full plate, but please note that you will be billed for every available position.
For our clients submitting in bulk we also offer partial services. BigDye 3.1 labeled fragments may be submitted for CleanSeq and Analysis. Purified samples ready for sequencing may be submitted for 3730xL Analysis Only.
For our clients submitting in bulk we also offer partial services. BigDye 3.1 labeled fragments may be submitted for CleanSeq and Analysis. Purified samples ready for sequencing may be submitted for 3730xL Analysis Only.
Pre-Sequencing Services
In addition to our basic sequencing service, the BPF also offers purification services for whole plate orders of PCR products and bacterial pellets. Clients must supply sequencing primers along with their unpurified samples.Standard Service (BigDye, Purification & Analysis)
Additional Services (PCR Clean, BigDye, Purification, & Analysis)
| Service | Quantity | Harvard | Non-Harvard/Academic | Commercial |
|---|---|---|---|---|
| PCRClean, Standard Bulk Order 96-Well (1 Plate) | 1 | $6.00 | $6.90 | $7.80 |
| PCRClean, Standard Bulk Order 384-Well (1 Plate or more) | 1+ | $3.50 | $4.03 | $4.56 |
| PCRClean, Standard Bulk Order 96-Well (2 Plates) | 2 | $5.50 | $6.33 | $7.16 |
| PCRClean, Standard Bulk Order 96-Well (3 Plates) | 3 | $5.00 | $5.75 | $6.50 |
| PCRClean, Standard Bulk Order 96-Well (4 or more Plates) | 4+ | $3.50 | $4.03 | $4.56 |
Partial Services for Bulk Orders
| Service | Quantity | Harvard | Non-Harvard/Academic | Commercial |
|---|---|---|---|---|
| SEQClean & Instrument Run (1 to 3 96-Well Plates) | 1-3 | $2.50 | $2.88 | $3.26 |
| SEQClean & Instrument Run (1 or more 384-Well Plates) | 1+ | $1.50 | $1.73 | $1.96 |
| SEQClean & Instrument Run (4 or more 96-Well Plates) | 4+ | $1.50 | $1.73 | $1.96 |
| 3730xL Instrument Run Only (1 to 3 96-Well Plates) | 1-3 | $1.50 | $1.73 | $1.96 |
| 3730xL Instrument Run Only (1 or more 384-Well Plates) | 1+ | $1.00 | $1.15 | $1.30 |
| 3730xL Instrument Run Only (4 or more 96-Well Plates) | 4+ | $1.00 | $1.15 | $1.30 |
Sequencing Supplies
| Company | Item Name | Quantity | Harvard | Non-Harvard/Academic | Commercial |
|---|---|---|---|---|---|
| USA Scientific BPF Sequencing Supplies | 0.5ml Non-coded V-bottom tubes - 8 Pack | 8 Pack | $0.31 | $0.36 | $0.41 |
| USA Scientific BPF Sequencing Supplies | 0.5ml Non-coded V-bottom tubes - 96 Pack | 96 Pack | $3.43 | $3.95 | $4.46 |
| USA Scientific BPF Sequencing Supplies | 8 Place Decapper | each | $5.95 | $6.85 | $7.74 |
| USA Scientific BPF Sequencing Supplies | Bag of 960 Tubes | each | $29.50 | $33.93 | $38.36 |
| USA Scientific BPF Sequencing Supplies | Box with insert | each | $1.40 | $1.61 | $1.82 |
| USA Scientific BPF Sequencing Supplies | Box with insert | each | $1.40 | $1.61 | $1.82 |
| USA Scientific BPF Sequencing Supplies | CapCluster - 8 Pack | 8 Pack | $0.35 | $0.40 | $0.45 |
| USA Scientific BPF Sequencing Supplies | CapCluster - 96 Pack | 96 Pack | $4.10 | $4.72 | $5.33 |
| USA Scientific BPF Sequencing Supplies | CapMat | each | $4.60 | $5.29 | $5.98 |
| USA Scientific BPF Sequencing Supplies | Custom 0.5 ml Tube - 8 Pack | 8 Pack | $0.25 | $0.29 | $0.32 |
| USA Scientific BPF Sequencing Supplies | Custom 0.5 ml Tube - 96 Pack | 96 Pack | $2.95 | $3.40 | $3.84 |
| USA Scientific BPF Sequencing Supplies | Individual Caps - 8 Pack | 8 Pack | $0.35 | $0.40 | $0.45 |
| USA Scientific BPF Sequencing Supplies | Individual Caps - 96 Pack | 96 Pack | $3.85 | $4.43 | $5.01 |
| USA Scientific BPF Sequencing Supplies | Lo-Bo Rack | each | $15.10 | $17.37 | $19.63 |
| USA Scientific BPF Sequencing Supplies | Sheet of tags | each | $2.30 | $2.65 | $2.99 |
| USA Scientific BPF Sequencing Supplies | Single Place Decapper | each | $5.00 | $5.75 | $6.50 |
| USA Scientific BPF Sequencing Supplies | Starter Kit | each | $9.95 | $11.45 | $12.94 |
| USA Scientific BPF Sequencing Supplies | TPE CapCluster Mats - mat of 96 | each | $5.31 | $6.11 | $6.91 |
| USA Scientific BPF Sequencing Supplies | TPE Capband-8 Mat - 96 caps | 96 Pack | $5.28 | $6.08 | $6.87 |
| USA Scientific BPF Sequencing Supplies | TPE Capband-8 strip - 8 caps | 8 Pack | $0.48 | $0.55 | $0.62 |
** Non BPF tubes are automatically upcharged $1.00 per sample for labor/handling and the correct tube.
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.