AccuStart II PCR Genotyping Kit
Features & Benefits
- Simplified, completely reagent-based system requires minimal pipetting skill.
- Premixed electrophretic mobility loading dye reduces chances for post-PCR cross contamination.
- Stabilized 2X PCR SuperMix enables convenient room-temperature setup and is unaffected by repetitive freeze-thaw (>20X).
- High-yielding, ultrapure modified Taq DNA polymerase delivers robust, reliable duplex assay performance.
- Stringent, ultrapure antibody hotstart ensures sensitive and specific target amplification.
AccuStart II PCR Genotyping Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.
Accustart II vs KAPA: Mouse tail snips
AccuStart II Results more specific
KAPA PCR mix requires more optimization to reduce non-specific amplification products in multiplex PCR reactions. Lanes: 1 = mouse 1, ZPBP1. 2 = mouse 2, ZPBP1. 3 = mouse 1, ZPBP1/p53. 4 = mouse 2, ZPBP1/p53. 5 = mouse 1, p53. 6 = mouse 2, p53
- Extracta® DNA Prep for PCR (95091-02)
- Extraction Reagent
- Stabilization Buffer
- AccuStart II GelTrack PCR SuperMix (95136-500)
- 2X concentrated SuperMix containing optimized concentrations of molecular-grade MgCl2, dNTP blend, AccuStart II Taq DNA Polymerase, reaction buffer, stabilizers and electrophoretic mobility dyes (4kb & 50bp).
- Extracta® DNA Prep for PCR (95091-02)
Storage & Handling
Remove AccuStart II GelTrack PCR SuperMix from the kit box and store separately at or below -20°C. Extracta reagents can be stored at room temperature
AccuStart II GelTrack PCR SuperMix is stable for 1 year when stored in a constant temperature freezer at or below -20°C. For convenience, it may be stored unfrozen at 4°C for up to 6 months. Repeated freezing and thawing does not impair reagent performance. Thaw completely, pulse vortex to mix and briefly spin down to collect tube contents before opening.
Related ResourcesProduct ManualsProduct FlyersSafety Data Sheets (SDS)PublicationsFzd4 Haploinsufficiency Delays Retinal Revascularization in the Mouse Model of Oxygen Induced Retinopathy11, PLOS ONE - 2016AbstractMutations in genes that code for components of the Norrin-FZD4 ligand-receptor complex cause the inherited childhood blinding disorder familial exudative vitreoretinopathy (FEVR). Statistical evidence from studies of patients at risk for the acquired disease retinopathy of prematurity (ROP) suggest that rare polymorphisms in these same genes increase the risk of developing severe ROP, implying that decreased Norrin-FZD4 activity predisposes patients to more severe ROP. To test this hypothesis, we measured the development and recovery of retinopathy in wild type and Fzd4 heterozygous mice in the absence or presence of ocular ischemic retinopathy (OIR) treatment. Avascular and total retinal vascular areas and patterning were determined, and vessel number and caliber were quantified. In room air, there was a small delay in retinal vascularization in Fzd4 heterozygous mice that resolved as mice reached maturity suggestive of a slight defect in retinal vascular development. Subsequent to OIR treatment there was no difference between wild type and Fzd4 heterozygous mice in the vaso-obliterated area following exposure to high oxygen. Importantly, after return of Fzd4 heterozygous mice to room air subsequent to OIR treatment, there was a substantial delay in retinal revascularization of the avascular area surrounding the optic nerve, as well as delayed vascularization toward the periphery of the retina. Our study demonstrates that a small decrease in Norrin-Fzd4 dependent retinal vascular development lengthens the period during which complications from OIR could occur.A novel mouse model for the identification of thioredoxin-1 protein interactionsMichelle L. Booze, Free Radical Biology and Medicine - 2016AbstractThiol switches are important regulators of cellular signaling and are coordinated by several redox enzyme systems including thioredoxins, peroxiredoxins, and glutathione. Thioredoxin-1 (Trx1), in particular, is an important signaling molecule not only in response to redox perturbations, but also in cellular growth, regulation of gene expression, and apoptosis. The active site of this enzyme is a highly conserved C-G-P-C motif and the redox mechanism of Trx1 is rapid which presents a challenge in determining specific substrates. Numerous in vitro approaches have identified Trx1-dependent thiol switches; however, these findings may not be physiologically relevant and little is known about Trx1 interactions in vivo. In order to identify Trx1 targets in vivo, we generated a transgenic mouse with inducible expression of a mutant Trx1 transgene to stabilize intermolecular disulfides with protein substrates. Expression of the Trx1 “substrate trap” transgene did not interfere with endogenous thioredoxin or glutathione systems in brain, heart, lung, liver, and kidney. Following immunoprecipitation and proteomic analysis, we identified 41 homeostatic Trx1 interactions in perinatal lung, including previously described Trx1 substrates such as members of the peroxiredoxin family and collapsin response mediator protein 2. Using perinatal hyperoxia as a model of oxidative injury, we found 17 oxygen-induced interactions which included several cytoskeletal proteins which may be important to alveolar development. The data herein validates this novel mouse model for identification of tissue- and cell-specific Trx1-dependent pathways that regulate physiological signals in response to redox perturbations.Intensified vmPFC surveillance over PTSS under perturbed microRNA-608/AChE interactionT. Lin, Translational Psychiatry - 2016AbstractTrauma causes variable risk of posttraumatic stress symptoms (PTSS) owing to yet-unknown genome–neuronal interactions. Here, we report co-intensified amygdala and ventromedial prefrontal cortex (vmPFC) emotional responses that may overcome PTSS in individuals with the single-nucleotide polymorphism (SNP) rs17228616 in the acetylcholinesterase (AChE) gene. We have recently shown that in individuals with the minor rs17228616 allele, this SNP interrupts AChE suppression by microRNA (miRNA)-608, leading to cortical elevation of brain AChE and reduced cortisol and the miRNA-608 target GABAergic modulator CDC42, all stress-associated. To examine whether this SNP has effects on PTSS and threat-related brain circuits, we exposed 76 healthy Israel Defense Forces soldiers who experienced chronic military stress to a functional magnetic resonance imaging task of emotional and neutral visual stimuli. Minor allele individuals predictably reacted to emotional stimuli by hyperactivated amygdala, a hallmark of PTSS and a predisposing factor of posttraumatic stress disorder (PTSD). Despite this, minor allele individuals showed no difference in PTSS levels. Mediation analyses indicated that the potentiated amygdala reactivity in minor allele soldiers promoted enhanced vmPFC recruitment that was associated with their limited PTSS. Furthermore, we found interrelated expression levels of several miRNA-608 targets including CD44, CDC42 and interleukin 6 in human amygdala samples (N=7). Our findings suggest that miRNA-608/AChE interaction is involved in the threat circuitry and PTSS and support a model where greater vmPFC regulatory activity compensates for amygdala hyperactivation in minor allele individuals to neutralize their PTSS susceptibility.Core Binding Factor-β Knockdown Alters Ovarian Gene Expression and Function in the Mouse - me.2015-1312Kalin Wilson, Original Research - 2016AbstractCore binding factor (CBF) is a heterodimeric transcription factor complex composed of a DNA binding subunit, one of three runt-related transcription factor (RUNX) factors, and a non-DNA binding subunit, CBF. CBF is critical for DNA binding and stability of the CBF transcription factor complex. In the ovary, the LH surge increases the expression of Runx1 and Runx2 in periovulatory follicles, implicating a role for CBFs in the periovulatory process. The present study investigated the functional significance of CBFs (RUNX1/CBF and RUNX2/CBF ) in the ovary by examining the ovarian phenotype of granulosa cell-specific CBF knockdown mice; CBF f/f * Cyp19 cre. The mutant female mice exhibited significant reductions in fertility, with smaller litter sizes, decreased progesterone during gestation, and fewer cumulus oocyte complexes collected after an induced superovulation. RNA sequencing and transcriptome assembly revealed altered expression of more than 200 mRNA transcripts in the granulosa cells of Cbfb knockdown mice after human chorionic gonadotropin stimulation in vitro. Among the affected transcripts are known regulators of ovulation and luteinization including Sfrp4,Sgk1,Lhcgr,Prlr,Wnt4, and Edn2 as well as many genes not yet characterized in the ovary. Cbf knockdown mice also exhibited decreased expression of key genes within the corpora lutea and morphological changes in the ovarian structure, including the presence of large antral follicles well into the luteal phase. Overall, these data suggest a rolefor CBFs as significant regulators of gene expression, ovulatory processes, and luteal development in the ovary.(Molecular Endocrinology 30: 733–747, 2016)Molecular adaptations in vasoactive systems during acute stroke in salt-induced hypertensionClick here to see all PublicationsNicole Ventura, Molecular and Cellular Biochemistry - 2014AbstractInvestigations regarding hypertension and dietary sodium- both factors that influence stroke risk- have previously been limited to using genetically disparate treatment and control groups- namely the stroke-prone- spontaneously hypertensive rat and Wistar-Kyoto rat. In this investigation- we have characterized and compared cerebral vasoactive system adaptations following stroke in genetically identical- salt-induced hypertensive- and normotensive control mice. Briefly- ANP+/- (C57BJ/6 Ã— SV129 background) mice were fed chow containing either 0.8 \% NaCl (NS) or 8.0 \% NaCl (HS) for 7 weeks. Transient cerebral ischemia was induced by middle cerebral artery occlusion (MCAO). Infarct volumes were measured 24-h post-reperfusion and the mRNA expression of five major vasoactive systems was characterized using qPCR. Along with previous publications- our data validate a salt-induced hypertensive state in ANP+/- mice fed HS chow as they displayed left ventricular hypertrophy- increased systolic blood pressure- and increased urinary sodium excretion. Following MCAO- mice fed HS exhibited larger infarct volumes than their dietary counterparts. In addition- significant up-regulation in Et-1 and Nos3 mRNA expression in response to salt and stroke suggests implications with increased cerebral damage in this group. In conclusion- our data demonstrate increased cerebral susceptibility to stroke in salt-induced hypertensive mice. More importantly- however- we have characterized a novel method of investigating hypertension and stroke with the use of genetically identical treatment and control groups. This is the first investigation in which genetic confounding variables have been eliminated.