Life Science/

Sanger Sequencing

V-mix

 

 

We mix DNA and primer, sequence, purify, load and analyse the samples. You receive the complete unedited sequence. Here read lengths of over 1100 bp are possible.

StarSEQ® can only offer the unbelievable price because of its high degree of automation during the whole sequencing process. Please follow the recommended primer and DNA concentration, since these are essential for the success of the sequencing reaction. Unsuccessful reactions will also be billed. Abbreviation and numbering on top of the lid of your sample tubes must be identical to order form!

 

  1. Login.
  2. Fill in the order form electronically.
  3. Submit the order form.
  4. Receive the completed order form as PDF by e-mail and popup window.
  5. Print order form and sign it.
  6. Label the reaction containers legibly on the lids with an indelible black pen.
  7. Please send your samples and primers in 1.5 ml tubes.
  8. Protect the reaction containers with a box or something similar and send together with order form in a padded envelope to:

 

StarSEQ GMBH, Johann-Joachim-Becher-Weg 30a, 55099 Mainz, Germany

For any questions or requests on your samples please contact us with your copmplete personal details, date of received results and the abbreviation of your samples under the following E-mail address:
v-mix@starseq.com

 

Recommendations:
DNA: dissolved in 10-15 µl water or TrisHCl (10mM), pH 7-8 (NO EDTA!!!). Minimum concentration of your plasmid DNA template must be 100 ng/µl!
Amounts: purified PCR product: 200 bp: >100 ng; 500 bp: >200 ng 1 kb: >400 ng /sequencing reaction
Plasmid DNA: <3,5 kb: 350 ng; 3,5 – 4,5 kb: 400 ng; 4,6 – 5,5 kb: 450 ng; 5,6 – 6,5 kb: 500 ng; 6,6 – 7,5 kb: 550 ng; 7,6 – 10 kb: 600 ng; > 10 kb: 700 ng /sequencing reaction
Cosmid DNA, PACs, BACs: >1 µg /sequencing reaction
Primer: 3 µl of a 10 µMolar primer solution for each rxn, standard primer available. You find the list here. For 48- or 96-well plates only one primer should be used for the whole plate. Or, another pair plate for primers should be provided.
Melting temperature: 55 – 58°C
Optimal length: 18 – 25mer

 

Primer base composition: G or C should be at the 3’ end, try to avoid more than 3 G’s or C’s at the end and 4 identical bases in a row.

Order form: be sure to fill in sample concentration. StarSEQ will not measure the DNA concentration of your samples. If sample concentration is not specified StarSEQ will take 100 ng/µl as a basis. For V-mix plate please provide sample concentration together with sample name separated with forward slash for each well.
Tube/Plate: 1.5 ml tubes (also for primers or standard 96 well plate sealed with PCR foil, aliminium foil or strips
Label: legible, with indelible black pen (e. g. Staedtler permanent lumocolor; Art. Nr. 318-9; EAN 40 07817 304563). Note your abbreviation on the lids and number them continuously. Please do not use special characters for the sample names like /[\}&%. Underscores and hyphen could be used.
Dispatch: Protect the tube with suitable firm boxes and send them in stable padded envelope.

*All quoted prices are without tax.

 

Sequencing results: sequencing data (.ab1 and .seq files) will be send to you by email and can also be downloaded directly from your account. The sequencing results are stored in your account for three months.

 

Hints for successful sequencing:

  • Sanger sequencing is not a PCR! Please add only one primer, otherwise you will receive a heterogeneous sequence.
  • DNA for sequencing should always be washed 2-3 times! It is generally not sufficient to wash the DNA only once.
  • Spin the columns during the washing steps for 2 minutes, 30 sec is not enough.
  • After the last washing step, the final spinning time should be 3-5 minutes.
  • Be sure that your centrifuge reaches the acceleration needed in g (not rpm). If the centrifuge does not reach this acceleration, you need to prolong the spinning time.
  • Before eluting the DNA place the column with open lid on top of the bench for 10-15 minutes to allow for evaporation of residual traces of EtOH. This step is very critical. The most common problem in Sanger sequencing is caused by EtOH contamination resulting in fast decreasing signals and short read lengths.
  • Do not elute DNA in the buffer that comes with the kit, try always to elute in molecular grade water.
  • Avoid overloaded columns during plasmid preparation or gel elution. Do not try to get as much DNA as possible out of the preps. This will lead to massive contamination from cell debris/gel in the eluted DNA. The output DNA may work in PCR, but the quality is too poor for sequencing.
  • Check the TM of the primer; it should be 52 – 58°C. Please note that sequencing conditions are different from most PCR conditions. The final concentration of salt in the sequencing reaction is 80 mM Tris, 2mM MgCI2. Please check the TM using these conditions.
48- or 96-well V-mix plate
5,16 €
(per Sequence Read)
batch from 20 V-mix samples and up
5,99 €
(per Sequence Read)
batch of 1-19 V-mix samples
6,20 €
(per Sequence Read)

Existing customers: Login to MyAccount or order

 

Registration: new customers please click here to register. After you have submitted your registration you will receive a confirmation email with a link to finalize the registration process. You will then receive another email with your login details. If you do not receive any email please check your spam folder.

 

U-mix user: please click here

 

Attention: Webshop and sanger sequencing orders need independent logins because of different security levels.

For V-mix samples we offer special services like “hot start” protocolls or adding agents to remove secondary structures. We also offer enzymatically clean up of PCR products prior to sequencing. Please inquiry your options and prices for this additional services.

V-MIX SANGER SEQUENCING PROMOTIONS

Cannot be combined with other promotions or discounts. Valid until 31.10.2022.

V-mix prepaid package: 50 samples
189€
(3,78* €/rxn)
V-mix prepaid package: 100 samples
357€
(3,57* €/rxn)
V-mix prepaid package: 250 samples
867,50€
(3,47* €/rxn)
V-mix prepaid package: 500 samples
1680€
(3,36* €/rxn)
V-mix prepaid package: 1000 samples
3250€
(3,25* €/rxn)

…or simply order by e-mail: info@starseq.com

Read pdf

*All quoted prices are without tax.

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