What is the price for sequencing?
$3.50-15.00/reaction. Price based on volume per order. Please call for a quote.
What payment methods are accepted?
We accept PO numbers as well as all major credit cards.
What is the turnaround time?
Sequencing turnaround is usually 24 hours. Orders that are picked up/arrive before 1PM should be posted by 10-noon the next day. Orders that arrive in the afternoon should be posted by noon to 2PM.
What read length will I get?
800-1000bp
How do I order sequencing online?
Click here http://www.retrogen.com/pdf/EnteringRetrieving.pdf to download the instruction for ordering online.
What tubes should I use?
Smaller orders should use 1.5mL epi tubes. Screw cap tubes are more time consuming so pop caps should be used. Tube strips while convenient are hard to label and keep track of so for large orders (24+ samples) use 96 well plates. We recommend using strip lids to cap plates as they prevent leakage and cross contamination – especially for plates shipped overnight.
How should I label the tubes?
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.
How do I send my samples?
San Diego customers can request a sample pickup and our courier will retrieve them.
Other’s can overnight them to:
Retrogen, Inc.
Sequencing Department
6645 Nancy Ridge Dr.
San Diego, CA 92121
We don’t recommend using Dry Ice as this greatly increases the shipping costs.
How do San Diego customers schedule a pickup?
You can call, email, or request a pickup through our website.
In order to keep orders best organized under the new system the preferred method for ordering sample pickups is through your LIMS account. Make sure that you first have entered you contact information in your user profile so we will know where to pickup your order. Under the sequencing request section there is an option to request sample pickups. Select the time period the sample will be ready, enter time when prompted, and click the “submit” button.
When do I need to have my samples ready?
Morning Pickup: Samples should be ready by 11AM for San Diego.
Afternoon Pickup: Samples should be ready by 3PM for San Diego.
Afternoon Pickup: Samples should be ready by 1pm for north counties.
Can I send my samples on a Saturday?
Retrogen is closed over the weekends so samples mailed on a Friday will not be delivered to us until Monday.
How much DNA template/primer do I need to send? What concentration should it be at?
Please click here http://www.retrogen.com/index/submission_template2.htm for detailed information.
Can Retrogen prep my DNA for me?
Yes. Please call for a quote.
What universal priming sites are there in my vector?
Contact the vendor who supplied the vector. If you email a text file with the sequence of the cloning region we can check it for binding sites for universal primers.
What primers can Retrogen provide?
The following universal primers are available at no extra charge:
Primer Name |
Sequence |
AOX1-For |
GAC TGG TTC CAA TTG ACA AGC |
AOX1-Rev |
GCA AAT GGC ATT CTG ACA TCC |
BGHrev |
TAG AAG GCA CAG TCG AGG |
CMV-F |
CGC AAA TGG GCG GTA GGC GTG |
DON1-F |
TCG CGT TAA CGC TAG CAT GGA TCT C |
DON2-R |
GTA ACA TCA GAG ATT TTG AGA CAC |
EGFP-C |
ATG GTC CTG CTG GAG TTC |
EGFP-N |
CGT CGC CGT CCA GCT CGA CCA G |
GLprimer1 |
TGT ATC TTA TGG TAC TGT AAC TG |
GLprimer2 |
CTT TAT GTT TTT GGC GTC TTC C |
M13-40 |
GTT TTC CCA GTC ACG AC |
M13F |
GTT TTC CCA GTC ACG ACG TTG TA |
M13F-20 |
GTA AAA CGA CGG CCA GT |
M13R |
CAG GAA ACA GCT ATG ACC |
Sp6 |
ATT TAG GTG ACA CTA TAG |
T3 |
ATT AAC CCT CAC TAA AGG GA |
T7 |
TAA TAC GAC TCA CTA TAG GG |
T7term |
GCT AGT TAT TGC TCA GCG G |
pBADf |
ATG CCA TAG CAT TTT TAT CC |
pBADr |
GAT TTA ATC TGT ATC AGG |
pGEX3 |
CCG GGA GCT GCA TGT GTC AGA GG |
pGEX5 |
GGG CTG GCA AGC CAC GTT TGG TG |
pTriplEx3 |
ACT CAC TAT AGG GCG AAT TG |
pTriplEx5 |
CTC GGG AAG CGC GCC ATT GTG TTG GT |
How do I design primers?
They are usually 18-25bp. Annealing temperature should be 50-65º, ideally 55-60 º. 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.
Can Retrogen make/design primers for me?
We can design and/or synthesize your primers for you. Call for a quote.
If Retrogen synthesizes my primer can it be used again for future sequencing orders?
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.
If Retrogen synthesizes my primer will it be sent to me after being used for the sequencing order?
We will send your primers upon request only. A shipping charge will apply.
Can I use the same primers I used to amplify my PCR product to sequence it?
Yes.
If I send primers and template DNA premixed how much of each should I use?
DNA |
DNA |
Primer |
Total Vol. |
2X DNA |
2X Primer |
2X Total Vol. |
PCR product DNA |
50ng |
10pmoles |
10uL |
100ng |
20pmoles |
20uL |
Single stranded plasmid |
250ng |
10pmoles |
10uL |
500ng |
20pmoles |
20uL |
Double stranded plasmid |
500ng |
10pmoles |
10uL |
1000ng |
20pmoles |
20uL |
BAC |
2000ng |
10pmoles |
10uL |
4000ng |
20pmoles |
20uL |
Example 1: If your PCR template and primer are at the concentration of 20ng/uL and 10pmol/uL, respectively, you need to add 2.5uL of PCR template and 1uL of primer to the 1.5mL epi tube, and add 6.5ul of H2O to bring the total volume of 10uL.
Example 2: If your Plasmid template and primer are at the concentration of 200ng/uL and 10pmol/uL, respectively, you need to add 2.5uL of PCR template and 1uL of primer to the 1.5mL epi tube, and add 6.5ul of H2O to bring the total volume of 10uL.
How long do you keep my DNA/Primers?
Samples are stored in our freezer for at least a month.
Can I get my DNA/Primers back?
Yes, but shipping charges will apply so we advise you aliquot a portion of your sample for us and keep the rest.
How close to the primer can I get sequence?
Sequence usually appears about 25-30 bases from the primer.
Can you sequence genomic DNA?
Directly sequencing genomic DNA is not very practical. Contact our technical support for help with your genomic sequencing projects.
Do you have special conditions for GC rich/2ndary structures/siRNA/etc.?
Yes. Be sure to note on your order any known issues so we can use conditions optimized for your sample.
My template is XX% GC, is that GC rich?
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.
What is FDA quality sequencing?
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.
How will I receive my data?
Your data will be posted to our website: http://sequencing.retrogen.com
You will be given login information and instructions for signing in and downloading your results.
How will I know when my data is ready?
You will receive an email notifying you when the results are ready.
How do I view the chromatograms?
You will need a program to view the chromatograms. You can find links to download a free program here: http://sequencing.retrogen.com/cgi-bin/dna/tools.cgi
Why did my reaction fail/give poor results?
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 email sequencing@retrogen.com
Can you repeat my reactions?
We may repeat a portion of an order in an attempt to troubleshoot or if we think the results can be improved. Call or email sequencing@retrogen.com to discuss any reactions you think should be repeated. Our full repeat policy can be found here: http://www.retrogen.com/index/seq_quality_assurance2.htm |
