Service and Ordering

  

We provide three kinds of cloning service.

 

1. Clone into pLEICS vectors(s) - choice of 136 vectors.


We will prepare PCR products from supplied templates and primers (if PCR fails we will perform one round of troubleshooting), and clone into pLEICS vector(s) of choice. 

User guide:

  1. Choose your expression system (E. coli, mammalian cells, baculovirus, Pichia Pastoris, Drosophila, Xenopus and cell-free systems, etc.)
  2. Choose your pLEICS vector(s) from Available Vectors
  3. Decide pLEICS vector family (Multiple vectors from one set of primers)
  4. Design PCR Primers or/and PCR-based mutagenesis primers ( see Primer Design Guide for details)

  5.  Order your primers and dissolve your primers in autoclaved water (concentration 1µg/µl preferred)

  6.  Submit your cloning request with a filled Cloning Request Form (DOWNLOAD) or/and Mutagenesis Cloning

      Request Form (DOWNLOAD), PCR primers and PCR template to PROTEX.

      Normally, orders received by Friday 4:00pm should be ready to return before 5:00pm next Friday

  7.  We will email you when we complete your clones. Then you will send your clones for sequencing and you can 

       find sequencing primer details at Vector Details (PNACL has all PROTEX pLEICS sequencing primers)

 

2. Clone into customer's vector(s) adapted for LIC method.


We will adapt the customer's vector(s) to LIC vector(s) with fast positive colony screen. The cost is £180.00 per vector and will take 6-8 weeks. 

You provide: 1. the full DNA sequence with features 2. vector map 3. enzyme digestion sites for cloning 4. vector plasmid DNA (concentration> 50ng/µl, 100µl) 5. a filled request form.

Cost for subsequent cloning is the same as for option 1 and normally, orders received by Friday 4:00pm should be ready to return before 5:00pm next Friday.

 

3. Clone into customer's vector(s).


You provide:1. the vector plasmid(s) (concentration> 50ng/µl, 100µl) 2. Vector sequence with feature 3. vector map, 4. PCR templates 5. your vector based PCR primers (for ligase free cloning method) 6. vector based colony PCR primers. 7. a filled request form. We then prepare vectors and PCR products from supplied templates and primers.

  

(For >10 clones, it is cheaper to adapt your vector(s) to LIC vector(s))

Normally, orders received by Friday 4:00pm should be ready to return before 5:00pm on the third Friday.


To order please:

 

1) For cloning DOWNLOAD order form and email to slm75@le.ac.uk

2) For mutagenesis DOWNLOAD order form and email to slm75@le.ac.uk

3) Please bring primers, templates and request form (pLEICS vectors are supplied by us) to HWB Rm. 1/40 or contact Sharon Munday (slm75@le.ac.uk)

 

We also provide protein expression test service

We now offer 1ml His-tagged protein expression trials in E. coli.flasks.jpg You supply the expression constructs and we will transform them into BL21 (DE3) or BL21 (DE3)-Rosetta, or you supply the E. coli expression strains containing your constructs (must be IPTG inducible). The protein expression test will be carried out in a 1ml culture in 2ml 96-well deep block, and the results will be presented as an SDS-Gel picture of fractions of total protein, soluble protein, Ni column flow-through, wash and elution. Currently, we can do expression trials at either 37° or 20° C. We can provide commonly used media - LB, TB and SOB but if necessary you can supply your specific media.

 

This service will run once every 3 weeks. Normally, for orders received by Friday 4:00pm the results should be ready before 5:00pm  next Friday.

 

To order please:

1) DOWNLOAD the order form and email to slm75@le.ac.uk

 2) Please bring samples and materials to HWB Rm. 1/40

 Any particular requirements please call Ila Patel, 0116 2525277 or email ip14@le.ac.uk.

 

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