Genome Engineering Unit offers:  

• Cloning of DNA constructs. 
• Whole plasmid sequencing (Illumina NGS sequencing) 

Vectors 

We have a variety of different backbones available for cloning, including vectors for protein expression in E. coli or in insect cells, vectors for human cell line transfections, lentivirus vectors, retrowirus vectors. You can also provide your own vector.  

Full list of backbones available can be found here. 

Inserts:  

Your insert can be amplified from a plasmid, cDNA or order as a synthetic gene. Mus musculus, Danio rerio, Caenorhabditis elegans and HEK293 cDNAs are available, you can also bring your own cDNA. We can modify the original insert by insertions, deletions, point mutations, adding or changing a tag.   

Prices:  

Internal IIMCB users can find the price list here 

External clients are requested to contact: request@crisprmice.eu  

Cloning methods 

We mainly use the following cloning methods: 

• Digestion with restriction enzymes and ligation with T4 DNA ligase 
• Sequence and ligation independent cloning (SLIC) 
• Gibson Assembly 

• Overlap extension PCR cloning  

After a successful cloning user receives:  

• Plasmid DNA (miniprep, at least 0,5 µg DNA) 
• Plasmid map 

• Sequencing results (chromatograms) 
• Bacteria (strain used for cloning) with your plasmid – upon request. 

Small aliquots of your construct is stored in our bank for one year in case you need to receive your construct again.