FESTIVE CLOSURE

Festive Closure

The service will close down over the festive period. We aim to process any sequencing samples arriving before or on the first post of Friday 22nd of December. Any sequencing samples arriving after that will be stored and processed on our return (3rd January). For other sample types please contact us.

Services | Maxipreps (+DNA Sequencing)

Falcon 2059 Minipreps

Preparing high quality plasmid DNA is essential for downstream molecular biology activities or other biochemical and cell biology work. Many of our customers will prefer to undertake this activity themselves for a variety of reasons. However, for customers who cannot do this, or simply just don’t wish to, we can help!

We offer a range of services that encompass all aspects of plasmid DNA preparation and allow customers to decide how much (or little) they wish us to do for them. These services include:

  • Plasmid transformation
  • Colony isolation and growth
  • Plasmid DNA purification

We can easily prepare your plasmid(s) on either a small (mini-prep), medium (midi-prep) or large (maxi-prep) scale to suit your downstream needs. We can also sequence the resultant plasmid DNA to assist you further.

We encourage you to review the information below prior to submission of your plasmid-prep so that you get the most out of our service and so that we can achieve the best quality plasmid DNA prep for you.

REGISTER SUBMIT NOW SUBMIT NOW (+DNA Seq)

Materials Required

What customers can provide:

We are happy to be flexible regarding receipt of materials to suit your needs!

  • Plasmid DNA - Please review our Complete Maxi-Prep Service below
  • Bacterial colonies on a plate - Please review the information provided below
  • Inoculated cultures to be grown - Please review the information provided below
  • Fully-grown Culture/Spun-down pellets - Please review the information provided below

Labelling Samples

  • When sending multiple maxi-preps, please provide your samples in tubes labeled sequentially from "1" onwards
  • As customers’ samples will be provided back in much smaller tubes than the culture tubes, excess text will hamper accurate transfer of sample name from one tube to another

 

E. coli Strains

  • We have found that DNA isolated from those E. coli strains in which the Endonuclease A has been mutated, results in purification of DNA that is more fully intact (non-degraded)
    • Please consider this should you choose to send us starter cultures/spun down bacterial pellets
  • Please note we only use the DH5alpha E. coli strain in-house

 

Media & Growth Considerations

  • When growing your cultures, please use only LB broth
  • Using too large a volume, an inappropriate growth media (ie. TB broth) or growing for too long results in a lot of bacteria but not much plasmid
    • Ultimately, excess bacterial cell material can clog up the clearing filter and leads to loss of your plasmid material
  • Media Volumes:
    • High Copy Plasmid: 150 ml LB
    • Low Copy Plasmid: 250 ml LB
  • Time Considerations: We have found that conditions that favour plasmid isolation include those where growth of the culture does not exceed 16 hours

 

Bacterial Colonies on a Plate

  • After you have transformed your plasmid(s) of interest, you can send us your plate(s) and our facility will:
    1. Select 1 colony per plasmid 
    2. Grow maxi-prep cultures
      • Please inform staff about antibiotic resistance for each plasmid
    3. Isolate DNA
    4. Quantitate DNA concentration

 

Inoculated Culture To Be Grown

  • Starter Culture: Please send your starter culture (5ml) together with details of the volume of culture and antibiotic to use
  • Antibiotic: As standard we have -
    • LB + Ampicillin
    • LB + Kanamycin
    • If you require a different antibiotic, we ask you to please grow your own culture and provide these to the DNA Sequencing and Services facility
  • Our facility can also isolate DNA directly from a culture you have begun growing

 

Fully Grown Culture/Spun-Down Pellets (Centrifuged Samples)

  • Our facility can also isolate DNA directly from a culture you have already grown from:
    • A fully grown culture
    • Spun-Down Pellet
  • To get the best results, ensure that you drain off all the culture medium, since not doing so reduces the quality of the plasmid DNA produced.

 

DNA Isolation

  • The preps we perform are Invitrogen Pure-Link high speed maxi-preps.
  • After prepping, the DNA is checked for quantity and quality on our nanodrop spectrophotometer and the concentration will be provided with the sample.

 

Complete Maxi-Prep Service

Should you wish for MRC PPU DNA Sequencing and services to take care of your maxi-prep from start to finish, our facility can easily do so. Please provide samples of DNA and we can take it from there! We don't stipulate an exact amount of DNA, but if you send us 20ul of your plasmid at 10ng/ul, that will be enough. We will:

  1. Transform your DNA
  2. Pick a single independent colony to grow a culture per plasmid
  3. Isolate the DNA for you
  4. Quanitate DNA concentration

 

Sequencing Your Maxiprep

Having Your Maxiprep Sequenced

It is often important to confirm the sequence of one's insert before embarking upon additional planned research endeavors in the lab. This can easily be accomplished by sequencing the DNA template which you have just had prepared by us. Simply click "Maxiprep + DNA Sequencing" above.

During the submission process, we will request additional information that will allow our facility to carry out this sequencing service for you including:

  • Information regarding your primer
    • We have a number of Standard Primers available
    • Or you can provide your own at the time that you submit your miniprep sample
  • Whether your template is known to be GC-Rich

Please contact us if anything is unclear!

Safety Considerations

Safety

  • As with all our services where we are accepting live bacterial material, please note that we can only accept samples that have been classified as biohazard containment level 1 according to the UK Genetically Modified Organisms (Contained Use) Regulations and ammendments.
  • For more information on these regulations, please see the HSE website.
  • We may require proof of containment level to be supplied.

 

Additional Information

Potential Yield of Plasmid DNA

  • Please note that the quantity of plasmid recovered from a maxi-prep is dependent on many factors and not just the size of the pellet or how cloudy the culture looks
  • Plasmid Copy Number Considerations
    • High Copy Plasmids
      • We have found that the yield for a high copy number plasmid can be around 1mg total DNA.
      •  In our experience, we have also found that isolation of DNA from maxi-preps of high copy plasmids grown in a volume of 250ml routinely overload the columns we use and can result in a low purity product thus we discourage customers from proceeding with their maxi-prep in this manner. 
    • Low Copy Plasmids
      • When using low copy plasmids, please ensure that you have identified the optimal strain to use for plasmid growth prior to submitting your cultures.   

Pricing

SERVICE Maxiprep Alone Maxiprep + Seq (Basic) Maxiprep + Seq (Including Quanti-It*) Maxiprep + Seq (Augmented Protocol^) Maxiprep + Seq (Including Quant-It* & Augmented Protocol^)
Maxiprep - Isolation of DNA from cell pellet £35.00 £40.00 £41.00 £40.50 £41.50
Maxiprep - Beginning with inoculated cultures £35.00 £40.00 £41.00 £40.50 £41.50
Maxiprep - Beginning with bacterial colonies £35.00 £40.00 £41.00 £40.50 £41.50
Maxiprep - Complete Service (Beginning with DNA to be transformed) £45.00 £50.00 £51.00 £50.50 £51.50

 

  • *Quant-It: We are happy to quantitate the concentration of your template for only £1
  • ^Augmented Protocol: If your template has known secondary structure or a high GC content, £0.50 will be applied per reaction to facilitate the sequencing reaction using an 'Augmented Protocol'

 

Do you routinely need a large number of maxipreps prepared and sequenced?

Why not contact our staff to learn about savings options?

Our pre-payment option will help you save more per reaction!

Primer

  • Standard primers offered by MRC PPU DNA Sequencing & Services are free!
  • Custom primer synthesis will reflect the size of the primer – the cost to the customer is £0.50 per base

Learn more about all our pricing and turnaround.

Please note that prices are exclusive of VAT, which customers outside The University of Dundee will be charged at the appropriate rate (currently 20%) on all products and that all sales are subject to MRC PPU DNA Sequencing and Services' Terms and Conditions.

Turnaround Time

Starting from an already grown culture, our facility can routinely turn samples around within one day of receipt. Additional time will be needed in the following cases:

  • Submission of Starter Culture - Routinely 2 day turnaround time due to additional required steps (amplification of bacteria/plasmid)
  • Submission of Plasmid DNA - At least 3 day turnaround time due to additional required steps (transformation, culture growth).

 

Shipping

Shipping Details

  • Labels
    • Please ensure that samples are clearly labelled so that we can identify them.
  • Packing Materials
    • Eppendorf Tubes (if sending plasmid DNA to be transformed)
      • Please also ensure that your samples are well protected and packaged to meet Post-Office or courier regulations
        • A "Jiffy" style padded envelope for DNA samples in plastic tubes works well
        • We discourage use of regular envelopes as in the past, we have received envelopes with tears and/or crushed tubes in them
    • Agar Plates (if sending colonies to selected for growth)
      • Please also ensure that your samples are well protected and packaged to meet Post-Office or courier regulations
    • E. coli Pellets
      • None needed if hand delivering (internal customers)
      • Packing in dry ice within a polystyrene insulated container is the only packaging method we can accept for items sent by courier

 

Postage Costs

  • Customers are responsible for paying all postage / carriage charges
    • For DNA samples or agar plates, normal first class post should be sufficient
      • As this does not always arrive as "next day post", we recommend that customers who have deadlines to meet send their samples by a guaranteed next day delivery service
  • Insufficient postage provided on a package will cause a delay in use receiving the package
    • If MRC PPU DNA Sequencing and Services is then charged upon receipt, please note that we will need to pass this cost onto the customer

 

Postal Address:

DNA Sequencing and Services
Medical Sciences Institute
School of Life Sciences
University of Dundee
Dundee
DD1 5EH
UK

Telephone:

(01382) 388019

Fax:

(01382) 388729

Email:

info@dnaseq.co.uk

Do you need any help? Please get in touch and we’ll be happy to lend a hand.