AECOM Sequencing Facility
Introduction

Sequencing is performed with an ABI 3730 DNA Analyzer.  The facility is prepared to meet the evolving needs of your lab.  Please contact David Reynolds (email) with any questions or special requests.
 
Sample Preparation
Plasmid DNA may be prepared for sequencing with a variety of methods and kits.  The quality of sequence data is closely related to the purity of the template DNA.  For best results with plasmids use a mini spin column which removes residual salts, proteins, residual detergents and residual RNA..

PCR products should be purified by column if the PCR reaction is specific with no artifact and a tight single product.  If artifact is present, gel purification with a column is recommended. 

Primers should be diluted to 3.2 pmol/µl or 1 pmol/µl in pure water.  The final amount in the sample should be 3.2 pmols.  Do not attempt to pipette less than 1 µl of primer or template.  Use a P-10 or P-2 when pipetting 1 to 2 µl.

 
Sample Submission
Samples can be submitted in two formats:

1.  Individual Samples: should be submitted in 0.2 ml strip tubes with individual, attached caps (USA Scientific, cat. no.1402-3900).  The sample submission form has instructions for mixing the template and primer.

2.  Plate Format:  Samples should be submitted according to these instructions to qualify for plate discount.. 

 
Per Sample Fee Schedule:
Plate Format (72 to 94 Samples)  $    5.00
Cancer Center & Human Genetics Program Members  $    7.00
AECOM Investigators  $    8.00
External Lab (Academic) $   10.00
Plate Format , External (Academic)  $    8.00
Printed Chromatograms(not available for plate format) Free
Standard Primers: list of available primers Free
Gel Quantitation Standards for PCR templates Free

 
Data Return and Analysis
Data Return: There are two options for data return

1.  Default Option:  The facility provides a printout of the sequence chromatogram and a text file (*.seq) via e-mail.

2.  The facility provides an electronic chromatogram (*.ab1) and a text file (*.seq) via reststop (public server) or ftp with e-mail notification.  If you prefer this option, write "ab1 file" on the Special Request line of the sample form.  Please note this is the default option for plate format submissions.  

Analysis: Links to free programs for viewing or analyzing chromatogram files (*.ab1)

Chromas: Viewer for Windows

Editview: Viewer for Mac

Traceviewer: Viewer for Mac and Windows

Softgenetics: Demos for aligning and mutation discovery

Aligner Demo: View and align on Mac OS X or Windows

Sequencher Demo: Viewer and restriction mapping program (Mac or Windows)

 
Contact
David Reynolds, Facility Director   e-mail  

Laboratory  e-mail  

Albert Einstein College of Medicine

Ullmann Building Room 713

1300 Morris Park Avenue

Bronx, NY 10461

P: (718) 430-2657