Sequencing FAQs

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Sequencing FAQ

We offer highly competitive prices. Discounts are available for large orders. Please call for a quote.

We accept PO numbers as well as all major credit cards.

Sequencing results are available the day after we receive your samples. Your results will be online by 10:00AM Monday through Friday and by 12:00PM on Saturdays. Additional turnaround time may be required if your reactions need to be repeated.

Click here http://www.retrogen.net/pdf/EnteringRetrieving.pdf to download the instructions for ordering online.

Smaller orders should use 1.5mL Eppendorf (epi) tubes. Screw-cap tubes are more time-consuming, so pop-caps should be used instead. Tube strips, while convenient, are hard to label and keep track of, so please use 96-well plates for large orders (24+ samples). We recommend using strip lids to cap plates, as they prevent leakage and cross contamination – especially for plates shipped overnight.

The label on the tubes should match the name listed on the order form. It is advisable to have names that are at least somewhat unique so they can be distinguished from other orders. Labeling the tops of the tubes is preferable but not required.

San Diego customers can request a sample pickup and our courier will retrieve them. Others can overnight them to:
Retrogen, Inc.
Sequencing Department
6645 Nancy Ridge Dr.
San Diego, CA 92121

We do not recommend using dry ice, as this greatly increases the shipping costs.

You can call, e-mail, or request a pickup through our LIMS System or our website at: http://www.retrogen.net/index/pickup_request2.htm

La Jolla / Sorrento Valley

Please have your pickup scheduled and have your samples in the appropriate pickup location by 11:00AM for the first pickup. We will pick up your samples between 11:00AM and 1:00PM. For the second pickup, have your samples ready by 3:00PM and we will pick them up between 3:00PM and 5:00PM.

North San Diego County (North of Hwy 56)

Please have your pickup scheduled, electronically or by telephone, by 1:00PM and have your samples in the appropriate pickup location by 1:30PM. We will pick up your samples between 1:30 and 2:30PM and you will receive your results the following morning as usual.

San Diego (South of Hwy 52)

Please have your pickup scheduled, electronically or by telephone, and have your samples in the appropriate pickup location by 1:30PM. We will pick up your samples between 1:30 and 3:30PM and you will receive your results the following morning.

Orange County

Please have your pickup scheduled, electronically or by telephone, by 2:00PM and have your samples in the appropriate pickup location by 3:00PM. We will pick up your samples around 3:00PM and you will receive your results the following morning as usual.

Los Angeles County

Please have your pickup scheduled, electronically or by telephone, by 12:00PM and have your samples in the appropriate pickup location by 1:00PM. We will pick up your samples around 1:00PM and you will receive your results the following morning as usual. Please note: Los Angeles pickup area is limited to south of Burbank and west of Azusa. We currently do not offer pickup service in Diamond Bar, Pomona, or Claremont areas.

Riverside/San Bernardino

Please have your pickup scheduled, electronically or by telephone, by 11:00AM and have your samples in the appropriate pickup location by 12:00PM. We will pick up your samples around 12:00PM and you will receive your results the following morning as usual.

Retrogen is closed on weekends, so samples mailed on Saturday will be delivered to us Monday.

Yes. Please call for a quote.

Contact the vendor who supplied the vector. If you e-mail us a text file with the sequence of the cloning region, we can check it for binding sites for universal primers.

PRIMER NAMESEQUENCE
AOX1-ForGAC TGG TTC CAA TTG ACA AGC
AOX1-RevGCA AAT GGC ATT CTG ACA TCC
BGHrevTAG AAG GCA CAG TCG AGG
CMV-FCGC AAA TGG GCG GTA GGC GTG
DON1-FTCG CGT TAA CGC TAG CAT GGA TCT C
DON2-RGTA ACA TCA GAG ATT TTG AGA CAC
EGFP-CATG GTC CTG CTG GAG TTC
EGFP-NCGT CGC CGT CCA GCT CGA CCA G
GLprimer1TGT ATC TTA TGG TAC TGT AAC TG
GLprimer2CTT TAT GTT TTT GGC GTC TTC C
M13-40GTT TTC CCA GTC ACG AC
M13FGTT TTC CCA GTC ACG ACG TTG TA
M13F-20GTA AAA CGA CGG CCA GT
M13RCAG GAA ACA GCT ATG ACC
Sp6ATT TAG GTG ACA CTA TAG
T3ATT AAC CCT CAC TAA AGG GA
T7TAA TAC GAC TCA CTA TAG GG
pBADfATG CCA TAG CAT TTT TAT CC
pBADrCCG GGA GCT GCA TGT GTC AGA GG
pGEX3ATG CCA TAG CAT TTT TAT CC
pGEX5GGG CTG GCA AGC CAC GTT TGG TG
pTriplEx3ACT CAC TAT AGG GCG AAT TG
pTriplEx5CTC GGG AAG CGC GCC ATT GTG TTG GT

The following universal primers are available at no extra charge:

They are usually 18-25bp. Annealing temperature should be 50-65º, ideally 55-60º. Primers that can self-hybridize should be avoided. Also, primers should be very site-specific to your binding site. Primers with homology to alternate sites, especially at their 3’ ends, can cause multiple signals.

Yes, we can design and/or synthesize your primers for you. Call for a quote.

Yes. If you plan on reusing primers synthesized by Retrogen, you should ask us for its tracking number. Referring to the primer by this number will make it easy for us to locate.

We will send your primers upon request only. A shipping charge will apply.

DNAAMOUNTPRIMERTOTAL VOL.2X DNA2X PRIMER2X TOTAL VOL.
PCR product DNA (per KB) 60ng 10pmol 6μL 120ng 20pmol 12μL
Single-stranded Plasmid150ng10pmol6μL300ng20pmol12μL
Double-stranded Plasmid300ng10pmol6μL600ng20pmol12μL
BAC2000ng10pmol6μL4000ng20pmol12μL

**We recommend 2x amount of DNA in case we need to repeat the reactions**

Example 1: If your 500bp PCR template and primer are at the concentration of 20ng/μL and 10pmol/μL, respectively, you need to add 3μL of PCR template and 2μL of primer to the 0.2mL PCR 8‐tube strip, and add 7μL of H2O to bring the total volume to 12μL.

Example 2: If your plasmid template and primer are at the concentration of 100ng/μL and 10pmol/μL, respectively, you need to add 6μL of plasmid template and 2μL of primer to the 0.2mL PCR 8‐tube strip, and add 4μL of H2O to bring the total volume to 12μL.

Samples are stored in our freezer for at least a month.

Yes, but shipping charges will apply, so we advise you aliquot a portion of your sample for us and keep the rest.

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Sequence usually appears about 25-30 bases from the primer.

Directly sequencing genomic DNA is not very practical. Contact our technical support for help with your genomic sequencing projects.

Yes. Be sure to note on your order any known issues, so we can use conditions optimized for your sample. An additional charge will apply. For special sequencing pricing, please contact our sales department.

Templates that are 70+% should be sequenced with our GC-rich conditions. Also, when sequencing through small regions (30-40bp) of very high GC content (80+%), our GC-rich conditions can improve the results.

It is designing additional primers for reactions to generate overlapping sequences so you can be confident that every base has been called with 100% accuracy.

Your data will be posted to our website: https://order.retrogen.com/login/ You will be given login information and instructions for signing in and downloading your results.

You will receive an e-mail notifying you when the results are ready.

You will need a program to view the chromatograms. You can find links to download a free program here:https://order.retrogen.com/login/

We have a troubleshooting guide that can aid you in determining why a reaction may have failed. If you still have questions, please call our technical support or e-mail sequencing@retrogen.net.

We may repeat a portion of an order in an attempt to troubleshoot, or if we think the results can be improved. Call or e-mail sequencing@retrogen.net to discuss any reactions you think should be repeated. Our full repeat policy can be found here: http://www.retrogen.net/index/seq_quality_assurance2.htm