The optimized plasmids enable improved efficiencies in making transgenics. They have been rigorously tested and are routinely used in our custom production of transgenics.
Your workflow can be made more efficient with our optimized plasmids. We offer 3 plasmids:
- sqt-1 marker plasmid (100ng/µL, 15µL volume)
- myo-2::GFP marker plasmid (15ng/µL, 15µL volume)
- hygR selection plasmid (100ng/µL, 15µL volume)
sqt-1 marker plasmid (pNU1228)
The sqt-1 plasmid encodes for a cuticle collagen expression in C elegans.
Composition is sqt-1p::SQT-1(genomic)::sqt-1utr.
A single point mutation (e1350) within the gene produces a morphological phenotype similar to Rol due to an improper cuticle formation.
Expression of SQT-1 is seen during L2, L3, and L4 larvae and adult stages, but not L1 or dauer larvae.
Figure 1. Plasmid map sqt-1 marker
This plasmid is suggested as a co-marker of injection efficiency in C. elegans and other nematodes. Successful injections will display a Rol phenotype in L2-L4 larvae and adult worms. In the injection mix, a final concentration is commonly used at 10 ng/µL in a 100ng/µL total DNA. The reagent is supplied as a 10X stock.
myo-2::GFP marker plasmid (pNU675)
The myo-2::GFP construct expresses GFP in the pharynx in C. elegans.
Composition is myo-2p::eGFP(3x-syntrons)::tbb-2utr.
The myo-2 promoter drives GFP exclusively in pharynx of the animal.
Figure 2. Plasmid myo-2::GFP marker
This plasmid can be used as as a co-marker of injection efficiency in C. elegans or other similar nematodes. Successful injections will display a bright green pharynx. In the injection mix, a final concentration of 1.5 ng/µL is commonly used in a 100ng/µl total DNA. The reagent is supplied as a 10X stock.
hygR selection plasmid (pNU1254)
The HygR plasmid encodes for hygromycin resistance, a useful selection tool, resulting in broad expression of hygromycin B phosphotransferase (HPT) in C. elegans tissues.
The HygR plasmid is commonly added into an injection mix at a final concentration of 10 ng/µL in a 100 ng/µL total DNA mix.
Sufficient accumulation of HPT expression occurs 24 hours post injection and survival starting two days post Hygromycin B exposure (0.25 mg/mL) indicates resistance.
By day five of post hygromycin exposure, only resistant animals are present.
Figure 3. Plasmid hygR selection
This HygR plasmid is a good selection tool for use in C. elegans or other similar nematodes. Successful injections will be resistant to Hygromycin B (0.25mg/mL) in plates. Addition of HygR plasmid into an injection mix is commonly used at a final concentration of 10 ng/µL in a 100ng/µL total DNA. The reagent is supplied as a 10X stock.