*** After over 30 years in business, Sequetech is closing its lab operations permanently and the last orders to be processed must be received by Thurs. 7-20-23. Our billing office remains open and your results will continue to be available for you to access. Thank you for your patronage and support over the years. ***
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Requirements

Our requirements for sequencing your DNA samples

Preferred Shipping Containers and Temperatures

Plasmids / PCR products (in plates)

Plasmid Sequencing in PlatesContainer:

For overnight shipping, cap strips are highly recommended. Adhesive sheets or foil sometimes leak. Clearly label all plates.

Temperature:

Dry Ice or Room Temperature. Please do not use wet ice.

Plasmids / PCR products (in tubes)

DNA Sequencing in PCR Tubes

Container:

Tubes should have a tight fitting cap. Clearly label all tubes. Please do not send PCR cap strip tubes.

Temperature:

Dry Ice or Room Temperature. Please do not use wet ice.

Liquid Culture / Glycerol Stocks

Sequencing DNA from Glycerol Stocks

Container:

Liquid Cultures or Glycerol Stocks for RCA should be shipped with tight fitting caps for samples sent in tubes. For plates, cap strips are highly recommended. Adhesive sheets or foil sometimes leak. Clearly label all plates.  We recommend that you wrap your shipment with bubble wrap to protect the contents.

Temperature:

Please ship at ROOM TEMPERATURE. Please do not use any type of ice.

Colonies that you picked from Agar plates

Container:

If picked into tubes, the tubes should have a tight fitting cap. If picked into plates, cap strips are highly recommended. Adhesive sheets or foil sometimes leak. We recommend that you wrap your shipment with bubble wrap to protect the contents. Please do not send PCR cap strip tubes.

Temperature:

Please ship at ROOM TEMPERATURE. Please do not use any type of ice.

Agar plates

DNA Sequencing from Colonies picked from Agar Plate

Container:

Agar plates can be shipped as is. They must be wrapped in parafilm. We recommend that you also pad your agar plate with bubble wrap when shipping.

Temperature:

Please ship at ROOM TEMPERATURE. Please do not use any type of ice.

Required Template/Primer Amounts and Concentrations

Requirements for Single Primer Extension
Template Requirements
Template Type Amount Concentration Notes
Plasmids 200ng/Primer Extension >50ng/ul Submitting extra will allow for re-runs, optimization and troubleshooting. Primers and templates are stored for at least 3 months. We strongly recommend confirming PCR products by gel analysis.
PCR (>800bp) 50ng/Primer Extension >10ng/ul
PCR(<800bp) 10ng/Primer Extension >2ng/ul

 

Primer Requirements
Amount Minimum
Concentration
Notes
4 pmole/primer extension 2 uM Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.

 

PREMIXED Sample Requirements
Template Type Amount of Template Amount of Primer Total Volume
Suspended in water is preferable.
Plasmid 500 ng 6 pmoles 10 µL
PCR (100bp-500bp) 10 ng 6 pmoles 10 µL
PCR (500bp-1000bp) 25 ng 6 pmoles 10 µL
PCR (>1000bp) 60 ng 6 pmoles 10 µL
Requirements for Primer Walking Service
Template Requirements
Template Type Service
(Click to download form)
Amount Concentration
Plasmids Single-Strand Coverage 3 ug per Kb >0.10ug/ul
Double-Strand Coverage 6 ug per Kb
PCR Product Single-Strand Coverage 0.3 ug per Kb >10ng/ul
Double-Strand Coverage 0.6 ug per Kb
Requirements for RCA (Rolling Circle Amplification)
Template Requirements
Template Type Instructions/Notes
Single Colonies on Agar Plates (Sequetech Picks the Colonies) After analyzing the sequences, mini-cultures of the clones of interest can be returned (by request.)
Single Colonies from Agar Plates (Customer Submits Picked Colonies) Cells should be picked into 10ul growth media supplemented with the appropriate antibiotic (please identify the antibiotic.) Make sure all tubes and plates are tightly sealed and do not leak. Please do not freeze.
Liquid Cultures and Glycerol Stocks We request 10ul of culture grown with the appropriate antibiotic (please identify the antibiotic.) Make sure all tubes and plates are tightly sealed and do not leak. Please do not freeze.
Partially Purified or Dilute Plasmid Preps Please supply 10ul of DNA at concentrations as low as 10pg/ul.

 

Primer Requirements
Amount Minimum
Concentration
Notes
4 pmole/primer extension 2 uM Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.
Requirements for BDX Sequencing

Template Requirements (BDX)

Template Type Amount Concentration Notes
Plasmids * 200ng/Primer Extension >50ng/ul Submitting extra will allow for re-runs, optimization and troubleshooting. Primers and templates are stored for at least 3 months. We strongly recomment confirming PCR products by gel analysis.
     
     

*Turbo PCR Sequencing is recommended for sequencing problematic PCR products.

 

Primer Requirements
Amount Minimum
Concentration
Notes
4 pmole/primer extension 2 uM Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.
Requirements for Turbo PCR Sequencing
Template Requirements
Template Type Amount Concentration Notes
PCR (>800bp) 50ng/Primer Extension >10ng/ul Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.
PCR(<800bp) 10ng/Primer Extension >2ng/ul

 

Primer Requirements
Amount Minimum
Concentration
Notes
4 pmole/primer extension 2 uM Submitting extra will allow for optimization and troubleshooting. Primers and templates are stored for at least 3 months.