33 Proteomics, Genomics & Microarrays


Complete Cpf1-Based CRISPR Genome Editing System Available


Integrated DNA Technologies (IDT) is the first genomics company to develop and bring to market a complete ribonucleoprotein-based Cpf1 CRISPR system. The Alt-R™ A.s. Cpf1 CRISPR System inherits the optimised, efficient, and cost-effective traits of IDT’s innovative Cas9-based system while taking advantage of Cpf1’s natural AT-rich target sequence preference and ability to make staggered cuts. In addition, IDT is launching an associated range of CRISPR support tools to expand experimental options and capabilities for molecular biology researchers. The new tools extend the ease-of-use and performance of IDT’s Alt-R system through options for fluorescent visualisation, enhanced nuclease transfection, and genome editing detection. Together, the new expanded Alt-R range breaks barriers to wider target spaces not addressable by Cas9 systems alone, and provides a level of flexibility in experimental design not previously possible.


IDT’s Alt-R System already overcomes the limitations of using sgRNAs in the ribonucleoprotein (RNP) complex by enhancing editing efficiency and lowering toxicity. Now, in developing a complementary Cpf1-based system, IDT has opened up options for targeting AT-rich sequences. The new system includes the Alt-R A.s. Cpf1 nuclease, containing two integrated nuclear localisation sites, which complexes as an RNP with a minimal 41-44 nucleotide Alt-R CRISPR-Cpf1 crRNA. The system requires no tracrRNA, reducing potential reagent costs and experimental complexity.


The expanded range of Alt-R support tools now includes the Alt-R CRISPR-Cas9 tracrRNA—5’ ATTO™ 550, and Alt-R Electroporation Enhancers for Cas9 and Cpf1. The former allows in vivo fluorescent visualisation of the RNP complex and FACS enrichment of transfected cells without affecting RNP functionality, while the enhancers improve the efficiency of transfection using the Nucleofector (Lonza) and Neon (Thermo) electroporation systems. These new tools enable researchers to improve the overall efficiency of their genome editing over traditional methods. In addition, the new Alt-R Genome Editing Detection Kit provides a fast, easy, and low-cost method for confirming editing events with a T7 Endonuclease I (T7EI)-based mismatch detection system.


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New Mixer Mill Adapters for Rapid and Effi cient Cell Disruption and Sample Homogenisation


Retsch’s mixer mill MM 400 is a true multi- purpose mill in the lab: it is used for mixing and homogenising powders and suspensions within seconds and is also ideally suited for the disruption of biological cells as well as DNA/RNA and protein recovery.


A new adapter now allows for simultaneous cell disruption of up to 8 x 30 ml cell suspension in 50 ml Falcon tubes with glass beads. The oscillation frequency of the mill of up to 30 Hz and the process time can be individually adjusted according to application requirements. Depending on the type of cell (yeast, bacteria, microalgae, filamentous fungi) this method is greatly superior to manual procedures as it allows to reproducibly disrupt up to 240 ml of cell suspension within 20 seconds to 7 minutes without noticeable temperature increase. Another new adapter for 4 x 30 ml wide mouth bottles allows for processing up to 8 samples for applications like washing bacteria off tissue samples.


The adapter for 5 ml Eppendorf tubes is used for the homogenisation of cell suspensions, tough secretions or tissue pieces with a volume up to 3 ml (incl. buffer) in single- use tubes in one working run, i. e. without previous division of the sample. The adapter accommodates 5 Eppendorf tubes and two adapters can be clamped into the mill; hence, it is possible to simultaneously process 10 x 3 ml of sample material.


This highly flexible mixer mill accepts a variety of grinding jars and adapters and is also used for the homogenisation of plants, soft cell tissue but also of harder sample materials.


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