Introduction

The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

Details

Specifications

Features	Specifications
Main Functions	Isolation up to 500µg endotoxin-free plasmid DNA from 25-50ml bacterial culture. Recommend for High copy vector, Low elution volume, High concentration
Applications	Cell transfection, animal injection, etc.
Purification method	Midi spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	High copy plasmid vector
Sample amount	25-50ml LB
Yield	10-500μg
Elution volume	≥0.1ml
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	250μg

Principle

The HiPure Plasmid procedure is based on alkaline lysis of bacterial cells followed by adsorption of DNA onto silica in the presence of high salt. The unique silica membrane used in the kit completely replaces glass or silica slurries for plasmid DNA minipreps. The procedure consists of 3 basic steps: Preparation and clearing of a bacterial lysate by alkaline method，then transfer the supernatant to column to bind DNA. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High purity – purified plasmid can be directly used in sequencing, enzyme digestion and PCR, etc.
• Fast – silica gel column purification is much faster than ion exchange
• High concentration – easily obtain concentrations >1000ng/μl
• Small amount large extraction – using a small amount of column to achieve a large extraction yield (500μg)
• Low endotoxin content – the obtained plasmid can be directly used in cell transfection and animal injection

Kit Contents

Contents	P123102	P123102B
Purification Times	50 Preps	50 Preps
RNase A	30 mg	30 mg
Buffer E1	150 ml	150 ml
Buffer E2	150 ml	150 ml
Buffer E3	150 ml	150 ml
Buffer E4	150 ml	150 ml
Buffer E5	150 ml	150 ml
Buffer PW2*	50 ml	50 ml
Elution Buffer	30 ml	30 ml
MaxPure EF Mini Column	50	50
2 ml Collection Tubes	50	50
Lysate Clear Midi Syringe	50	50
Extend Tubes	50	50
50ml Centrifuge Tubes		50
Support Tubes		50

Storage and Stability

The kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve. After addition of RNase A and optional LyseBlue reagent, Buffer P1 is stable for 6 months when stored at 2–8°C.

Purchase Guide

Name	CAT NO	Features	Sampleamount	Elutionvolume	Yield
HiPure Plasmid Mini Kit	P1001	Classic, Regular application	1-5ml LB	30-50μl	5–35µg/5ml
HiPure Plasmid Mini Kit II	P1002	Classic, Regular application	5-15ml LB	75-100μl	20-80µg/15ml
HiPure Plasmid Plus 96 Kit	P1006	Classic, Regular application	1-1.5ml LB	70-150μl	1-15μg/1ml
HiPure Fastfilter Plasmid Midi Kit	P1013	Fast	30-50ml LB	0.3-0.8ml	50–250µg/50ml
HiPure Fastfilter Plasmid Maxi Kit	P1014	Fast	100-200ml LB	0.5-1.5ml	0.4–1mg/200ml
Low endotoxin/Endotoxin-free Plasmid
HiPure Plasmid EF Maxi Kit	P1156	Classic method, General for all kinds of vectors	100-200ml LB	≥0.7ml	200–1500µg
HiPure Plasmid EF Mini Kit	P1154	Recommend for low copy vector, Thoroughly remove RNA	5-15ml LB	≥60μl	10–80µg
HiPure Plasmid EF 96 Kit	P1157	Recommend for low copy vector, Thoroughly remove RNA	1-5ml LBx96	≥75μl	30µg
HiPure Plasmid EF Midi Kit	P1231	Recommend for High copy vector, Low elution volume, High concentration	25-50ml LB	≥100μl	10-500μg
HiPure Plasmid EF Mega Kit	P1116	Recommend for High copy vector(>3μg/ml)	500ml LB	≥2ml	1-10mg

For any technical problems or customized products, please contact us.

F&Q about Endotoxin-free Plasmid Extraction Kit — P1156 ←click here

The HiPure Midi system provides a fast, simple, and cost-effective plasmid DNA midiprep method for routine molecular biology laboratory applications. HiPure Midiprep Kits use silica membrane technology to eliminate the cumbersome steps associated with loose resins or slurries. Plasmid DNA purified with Mini Kits is immediately ready for use. Phenol extraction and ethanol precipitation are not required, and high quality plasmid DNA is eluted in a small volume of Tris buffer or water.

• Screening of Microcystins in water samples at 6 ppb or lower
• Format: 10 tests (5 tests/5 control)
• Water Sample Bottles
• Negative Control
• 1mL Syringe
• Sample Dilution Buffer also sold separately
• Sample Filters

Description

Attogene’s Microcystin Test Kit (Rapid – Recreational Water) can be used to detect microcystins in liquid samples; highly sensitive, rapid, robust screening kit for algae toxins microcystins and nodularins.

The most frequently reported cyanobacterial toxins are the hepatotoxic microcystins (MCs). MCs are peptides with a molecular weight ranging from 900 to 1,100 Da. They consist of seven amino acids of which the two terminal amino acids of the linear peptide are condensed to form a cyclic compound.

A tiered notification system which takes different actions based on different numeric thresholds for microcytin-LR concentrations in recreational waters has been developed. This is guidance that allows states to take various actions—such as posting information about harmful algal blooms (HABs), issuing a recreational public health advisory, or temporarily closing recreational waters through a no contact advisory—depending on the severity of the bloom event.

Microcystin-LR

Informational sign postings about HABs at recreational waters: < 6 μg/L
Recreational public health advisory: 6 μg/L
Elevated recreational public health advisory (e.g. no contact): 20 μg/L

Harmful Algae Bloom – NOAA

Screening of Microcystins in water samples at 6 ppb or lower
Format: 10 tests (5 tests/5 control)
Water Sample Bottles
Negative Control
1mL Syringe
Sample Dilution Buffer also sold separately
Sample Filters

• HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® Taq DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.HiDi® Taq DNA polymerase harbours a nuclease function and therefor is also suitable for use with hydrolysis probes (TaqMan® probes etc.). It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control and mutation identification in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® Taq DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)
  For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.

HiDi is available as:

HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)

Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.

Example Results – There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.

HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).

Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.