Core/facility: Keck DNA Sequencing Facility Offering: Sanger sequencing

Plate sequencing

Plate-based sequencing is a cost-effective and time-saving service recommended for those experienced in their methodology for template and primer preparation.

Minimum charge for this service is 48 samples, but you may submit fewer. Overall, it is a cheaper and faster way to get your samples sequenced. We sequence and charge for all samples submitted on the plate, regardless of the results, unless it is a clear technical error on our part.

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Sanger sequencing template prep instructions

2uL 4uM Primer + DNA(see table) + (sterile H2O, if necessary) = 18uL

Product type	Product size	DNA (ng)	Ordering notes
PCR	100-300 bp	40-50	*Can be ordered with Difficult Template or shRNA protocol
	300-500 bp	40-160
	500-1000 bp	500
Plasmid		1000-1500
Single-stranded M13 or phagemic DNA		200-300	Order as single-stranded
BAC and phage lambda DNA		4000	Order as BAC

Each reaction should include 2uL of a 4uM Primer. One primer per reaction.
The total volume should be 18uL. If extra volume needed, use sterile H2O.

Submission requirements

Each well of your plate should contain (1) primer and (1) template.

Semi-skirted plates — 2 mL semi-skirted plates

Plate specifications

All 96-well plate submissions must be submitted in semi-skirted, 2 mL 96-well plates with clear wells, and sealed with strip caps or sealing film/foil. For more acceptable plate types, please visit our Plasticware requirements page.

Fully skirted, non-transparent wells — Incorrect fully-skirted, non-transparent wells

Incorrect plate type

This plate is an example of an incorrect submission. It is fully skirted. It also has opaque wells. We need the wells to be transparent so we can verify that sample is present, and it helps us to verify the processing is correct.

96-well Plate with strip caps — Plate sealed with strip caps

Labeling and sealing instructions

Be sure to label your submission with your researcher code and your unique plate name.

Cap the plate firmly using the strip caps or sealing film and wrap each plate with parafilm. (Sealing film is preferable to strip caps.)

Plate filled by rows — Plate correctly filled A01-A12, B01-B12, etc.

Correct plate setup

When submitting fewer than 96 samples on a plate, please submit the samples in consecutive rows. Begin filling the wells at A1 through A12, then B1-B12, C1-12, etc.. If there are any empty wells in the middle of your submission, you will be charged for them.

Incorrect plate setup

This plate is filled incorrectly; it has missing wells and does not run from well 1-12 in a row.

Confirmation Page — Order confirmation sheet

Confirmation sheet

Plate submissions should be dropped off with an order confirmation sheet (available to print after web ordering via our “Sample submission” page).

Sample sheet template — Optional sample sheet format

[Optional] sample sheet

You may submit a sample sheet with your submission if you wish. If you do not send us an Excel form with your sample names, they will be named as follows: [Researcher code]-[Plate name]-[well position].