Introduction

This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.

Details

Specifications

Features	Specifications
Main Functions	Isolation gDNA from biological sample and remove host DNA
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Culture medium, swab, parasitic blood, tissue, sputum, etc.
Sample amount	Blood sample, homogenate, plasma, brain effusion, swab immersion solution, centrifuged concentrated liquid, etc.:0.5-1ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica Column purification. This product efficiently depletes human and animal host DNA and yields enriched bacterial DNA. An optimized combination of mechanical and chemical lysis allows efficient disruption of bacterial cells. Target DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High concentration – the host DNA is digested by dnase, without contamination of host DNA
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time
• Wide applicability – it can be used in blood, tissue, intestinal contents and other samples

Kit Contents

Contents	D314802	D314803
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns I	50	2 x 125
2ml Collection Tubes	50	2 x 125
2ml beads Tubes	50	250
Buffer DRB	15 ml	60 ml
Buffer ES	6 ml	30 ml
Reagent DX	0.5 ml	1 ml
Buffer DL	30 ml	120 ml
Buffer GW1	22 ml	110 ml
Buffer GW2	12 ml	50 ml
DNase I (Powder)	6 mg	30 mg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer AE	15 ml	60 ml

Storage and Stability

DNase I and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.

Description 

The GetClone™ PCR Cloning Vector II is a positive selection system for high efficiency cloning of blunt end DNA or amplicons. This cloning vector contains a lethal gene which can be disrupted by ligation of a blunt end DNA insert into the cloning site. Only colonies with inserted vectors are able to propagate, eliminating the additional needs of IPTG and X-Gal for blue/white screening. This cloning vector includes ampicillin and kanamycin resistance genes that can meet the needs of most users. 

Features

• Cloning efficiency greater than 90% 
• IPTG and X-Gal are not required 
• Accepts a wide range of insert/vector ratios 0.5:1 to 12:1 
• Accepts insert size from 6 bp to 11 kb
• The phosphorylation of PCR fragments is not required
• Accepts blunt end amplicon or DNA fragment (not for sticky ends)
• Ampicillin and kanamycin selection markers

Storage

-20°C for 24 months

The GetClone™ PCR Cloning Vector II is a positive selection system for high efficiency cloning of blunt end DNA or amplicons. This cloning vector contains a lethal gene which can be disrupted by ligation of a blunt end DNA insert into the cloning site. Only colonies with inserted vectors are able to propagate, eliminating the additional needs of IPTG and X-Gal for blue/white screening. This cloning vector includes ampicillin and kanamycin resistance genes that can meet the needs of most users.

endo-BCN-PEG4-Biotin is a biotinylated PEG linker containing a BCN moiety for click chemistry and a PEG4 arm that imparts water solubility for the biotinylated molecule.

endo-BCN-PEG4-Biotin is a biotinylated PEG linker containing a BCN moiety for click chemistry and a PEG4 arm that imparts water solubility for the biotinylated molecule.