Zyppy-96 Plasmid Kit

D4041 / D4042 / D4043


Zyppy-96 Plasmid Kit

D4041 / D4042 / D4043

Cat # Name Size Price Quantity
D4041 Zyppy-96 Plasmid Kit 2 x 96 Preps $370.00
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D4042 Zyppy-96 Plasmid Kit 4 x 96 Preps $662.00
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D4043 Zyppy-96 Plasmid Kit 8 x 96 Preps $1,190.00
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Documents


Protocol: PDF   |   SDS (MSDS): PDF   |  
Pellet-free, high-quality plasmid DNA in only 8 minutes

Highlights


  • Pellet-Free: Direct lysis procedure omits cell-pelleting and resuspension steps.
  • High Quality: Ready for PCR, sequencing, cloning, and transfection.
  • Streamlined Protocol: Culture, lyse, and neutralize in the same plate, saving time and reducing plastic waste.

Description


The Zyppy-96 Plasmid Miniprep is the fastest and simplest high-throughput plasmid DNA kit available for efficient isolation of plasmid DNA from E. coli. This plasmid DNA kit features a modified alkaline lysis system that is pellet-free. This completely bypasses tedious centrifugation, pelleting, and re-suspension steps common to conventional procedures. Instead, the uniquely formulated Deep Blue Lysis Buffer is added directly to the bacterial cultures in a 96-well block. Buffer neutralization and lysate separation steps are assisted with a specially designed Neutralization/Clearing Buffer. Eluted plasmid DNA is high quality, endotoxin-free, and well-suited for use in restriction endonuclease digestion, DNA ligation, PCR, transcription, sequencing, and other sensitive downstream applications including transfection.

Technical Specifications


Applicable For Ligation, sequencing, restriction endonuclease digestion, in vitro transcription, and other sensitive applications requiring pure DNA.
Elution Volume ≥ 30 µl
Equipment Centrifuge with micro plate carriers
Processing Time 45 min
Purity Typical Abs 260/280 ≥ 1.8.
Size Range Up to 25 kb
Yield Up to 10 µg per preparation depending on the plasmid copy number, culture growth conditions, and strain of E. coli processed.

Product FAQ


Q1: Can E.coli grown in enriched growth media be used with this kit?

Q2: What is the composition of the Zyppy Elution buffer?

Q3: Can the purified plasmid be transfected into eukaryotic cell lines?

Q4: Can the Zyppy Plasmid kits be used to isolate large constructs (BAC/PAC)?

Q5: What are the endotoxin levels in plasmid DNA isolated with the Zyppy Kits?

Q6: I accidentally left my Neutralization Buffer at room temperature. Will it be okay?

Q7: I ran out of Zyppy Wash Buffer. Can I substitute it with a homemade solution or Wash Buffer from another kit?

Citations


GPR139 is a Gq‐coupled receptor activated by the essential amino acids L‐tryptophan (L‐Trp) and L‐phenylalanine (L‐Phe). We carried out mutagenesis studies of the human GPR139 receptor to identify the critical structural motifs required for GPR139 activation. We applied site‐directed and high throughput random mutagenesis approaches using a double addition normalization strategy to identify novel GPR139 sequences coding receptors that have altered sensitivity to endogenous ligands. This approach resulted in GPR139 clones with gain‐of‐function, reduction‐of‐function or loss‐of‐function mutations. The agonist pharmacology of these mutant receptors was characterized and compared to wild‐type receptor using calcium mobilization, radioligand binding, and protein expression assays. The structure‐activity data were incorporated into a homology model which highlights that many of the gain‐of‐function mutations are either in or immediately adjacent to the purported orthosteric ligand binding site, whereas the loss‐of‐function mutations were largely in the intracellular G‐protein binding area or were disrupters of the helix integrity. There were also some reduction‐of‐function mutations in the orthosteric ligand binding site. These findings may not only facilitate the rational design of novel agonists and antagonists of GPR139, but also may guide the design of transgenic animal models to study the physiological function of GPR139.

Wang, Lien, et al. "Mutagenesis of GPR139 reveals ways to create gain or loss of function receptors." Pharmacology Research & Perspectives 7.1 (2019): e00466.

Multiple sclerosis (MS) is a neurodegenerative disease of the central nervous system (CNS). Minimally invasive biomarkers of MS are required for disease diagnosis and treatment. Differentially methylated circulating-free DNA (cfDNA) is a useful biomarker for disease diagnosis and prognosis, and may offer to be a viable approach for understanding MS. Here, methylation-specific primers and quantitative real-time PCR were used to study methylationpatterns of the myelin oligodendrocyte glycoprotein (MOG) gene, which is expressed primarily in myelin-producing oligodendrocytes (ODCs). MOG-DNA was demethylated in O4+ ODCs in mice and in DNA from human oligodendrocyte precursor cells (OPCs) when compared with other cell types. In the cuprizone-fed mouse model of demyelination, ODC derived demethylated MOG cfDNA was increased in serum and was associated with tissue-wide demyelination, demonstrating the utility of demethylated MOG cfDNA as a biomarker of ODC death. Collected sera from patients with active (symptomatic) relapsing-remitting MS (RRMS) demonstrated a higher signature of demethylated MOG cfDNA when compared with patients with inactive disease and healthy controls. Taken together, these results offer a minimally invasive approach to measuring ODC death in the blood of MS patients that may be used to monitor disease progression.

Olsen, John A., et al. "A Minimally-invasive blood-derived biomarker of oligodendrocyte cell-loss in multiple sclerosis." EBioMedicine 10 (2016): 227-235.

Kit Components