Bulk Sequencing: (48 or more Pre-Defined samples in a 96-well plate)

To receive a discount for submitting 48 or more Pre-Defined samples, please submit your Pre-Defined samples in a 96-well PCR plate using the column or vertical loading scheme shown above for Bulk Sequencing of Pre-Mixed samples. If you are requesting the use of 10 or fewer primers, then submit them separately in labeled tubes. If you are requesting the use of more than 10 primers, then they must be submitted separately in a 96-well plate arrayed to match with the appropriate template.

Custom DNA Sequencing (Additonal charges may apply):

GENEWIZ will adjust the DNA concentration and add your separately submitted primer, or if you specify, a free GENEWIZ Universal Vector Primer.

• Submit your DNA in an 8-strip 0.2 ml PCR tube (please feel free to use 1.5 ml eppendorf tubes if you are submitting one DNA sample with many primers). Make sure that your tubes are clearly marked with either the tube label or the sample name.
• Please submit 500 -800 ng or more of plasmid DNA, or 80-150 ng or more of PCR fragment for each sequencing reaction.
• Sequencing primer (5 µl per reaction) should be submitted as 5 pmol/µl (5 µM) in dH₂0. The primer can be submitted in 0.5 ml or 1.5 ml eppendorf tubes with the primer name and concentration clearly labeled.

PCR Clean-up

If you have a single PCR band and you would like GENEWIZ to perform the PCR clean up, you can select Custom Sequencing and choose PCR Clean-up from the Special Protocol column. We also request that you submit a gel image of your PCR product with the amount loaded in each well clearly labeled.

If you would like to perform the PCR clean up, we highly recommend using the ExoSAP-IT kit from USB Corporation (Cat. #78200).

Sequencing from Bacterial Colonies (Additional charges apply):

• Submit your bacterial colonies on an agar plate. Securely wrap the plate with parafilm for transport and send at room temperature. Be sure to avoid shipping on Friday so that the colonies do not overgrow.
• Sequencing primer (5 µl per reaction) should be submitted as 5 pmol/µl (5µM) in dH₂0 in a 1.5 or 0.5 ml eppendorf tube.
• If you would like us to pick specific colonies, be sure to clearly circle or mark the colonies you need sequenced. If you just need us to randomly pick colonies, please clearly identify the number you would like us to sequence per agar plate.

Sequencing from Glycerol Stock (Additional charges apply):

• Submit your bacterial cultures with 8-30% glycerol in 1.5 ml or 2 ml eppendorf tubes. We need 10-50 µl per reaction. All glycerol stock samples must be sent to us on dry ice, so please avoid sending on a Friday.
• Sequencing primer (5 µl per reaction) should be submitted as 5 pmol/µl (5µM) in dH₂0 in a 1.5 or 0.5 ml eppendorf tube.

Bulk Sequencing from Bacterial Colonies or Glycerol Stock: (48 or more samples on an arrayed agar plate or in a 96-well plate)

Bacterial Colonies - The preferred colony submission method and best way to prevent cross contamination and sample mix-ups is by growing colonies in a grid pattern on an arrayed agar plate. This is an especially good strategy for colonies used in downstream applications, such as plasmid DNA preparation. We also bulk sequence colonies that are randomly distributed on an agar plate; however, these colonies will not be labeled unless specifically requested in the DNA sequencing form. For example, if there is a possibility that a colony will be used for DNA preparation after it has been sequenced, then write "May need DNA prep service after sequencing- please label colonies" in the "Notes" of the DNA sequencing order form.

Agar plates can be preprinted using a "stamper" for creating an evenly spaced grid of bacterial colonies. Alternatively, you can draw a grid on the back of the plate with alcohol resistant indelible ink.

Shipping: To ship the plates to GENEWIZ, be sure to secure the lid to the plate with parafilm and ship it with some padding. We also highly recommend using a box instead of an envelope to help prevent damage during transport.

Glycerol Stock- Any 96-well PCR plate or V-bottom 96-well plate can be used. The plates need to be sent on dry ice. Please provide 30-50 µl of your 8-30% glycerol stock for each reaction.

Sequencing from Phage Supernatant (Additional charges apply):

• Please submit 50µl of your phage supernatant in 8-strip PCR tubes or 1.5 ml eppendorf tubes that are clearly labeled with the sample name.
• Indicate on the submission form the phage titer. This can be put in the comments section or hand-written on the final printout that will be sent in with your samples.
• The primer should be supplied at 5 pmol/µl in a separate tube clearly labeled with the primer name and concentration.
• Please allow us to perform a small pilot study consisting of 3-4 dilutions that will help determine the optimal sequencing conditions.

BACend Sequencing (Additional charges apply):

Please be aware that due to the difficult nature of this type of sequencing, we suggest that you send a small pilot study. GENEWIZ asks that the DNA and primers be supplied in concentrated form so that we can use more DNA and primer to increase the chance of success.

• Please submit 2 µg or more of your BACend DNA (100k ?200k) for each sequencing reaction. The DNA should be at least 150 ng/µl or more and you should submit 15µl per reaction. Clearly label the DNA tube with the sample name.
• Primers should be submitted in separate tubes at a concentration of 10 pmol/µl (10 µM) or more ?please provide 5 µl of primer for each sequencing reaction. Also make sure that the primer is clearly labeled with the primer name and concentration.

Primer Walking:

To order primer walking services, please download Primer Walking Service Order Form and email your order to dnaseq@genewiz.com. Also print out a hard copy of the order form and send in with your samples.

Plasmid: Submit 1 µg per kb of target sequence for single stranded coverage and 2 µg per kb sequence for double stranded coverage. Submit at least 10 µg plasmid if region of interest is of unknown length.

Primers: GENEWIZ can synthesize initial primer for you if the primer sequence is provided. If you have primers that can be used in the initial round of sequencing, please submit at least 10 µl each at 5 µM.

Packaging: GENEWIZ recommends 1.5-ml screw cap tubes for DNA and primer submission. Please label the tubes clearly with sample/primer name and concentration. Tighten the tube caps and use enough cushion in the package. Please ship your package to GENEWIZ with standard overnight service or contact us at dnaseq@genewiz.com to find out if there is a GENEWIZ sample drop box near your location.