CRISPR/Cas9 - Services Offered by Microsynth
- Synthesis of tracrRNA and crRNA oligonucleotides1
- Synthesis of DNA oligonucleotides for molecular cloning of the target-specific region of your gRNA
- Sanger sequencing to verify the sequence of your CRISPR plasmid
- Analysis of specific CRISPR/CAS9 edited clones by PCR, cloning and Sanger sequencing
- Next generation sequencing to validate single CRIPSR/Cas9 targets by using an amplicon deep-sequencing approach
- Measurement of expression levels of targeted genes using a real-time PCR approach including assay development
- Detection of the edits in CRISPR/Cas9 screenings. Microsynth has developed high-input protocols for DNA isolation, PCR and sequencing to address the complexity of a CRISPR/Cas9 screening
In many projects Microsynth has gained experience to choose the right method for a specific analysis. Please just contact us for a detailed project discussion.
1 The benefits of using two synthetic RNAs (tracrRNA and crRNA) compared to a single guide RNA (sgRNA) expressed from a vector is that the multiple crRNAs can be ordered and transfected within a few days. This allows for rapid screening of several target sites within a single gene or knocking out several genes quickly, presuming your cells are transfectable. Expressed sgRNAs require individual cloning of each target sequence into an expression plasmid, growing and selecting several clones to control by sequencing, then prepping and purifying DNA suitable for transfection. This is a very time-consuming approach when you wish to test several crRNA candidates per gene or would like to knockout multiple genes quickly.