Biotin-PEG12-DBCO is a biotinylating reagent comprised of DBCO, a click chemistry handle that is frequently used to conjugate with azides on target molecules, and biotin, which is useful for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins. The PEG12 imparts the hydrophilicity of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Biotin-PEG12-DBCO is a biotinylating reagent comprised of DBCO, a click chemistry handle that is frequently used to conjugate with azides on target molecules, and biotin, which is useful for affinity-based applications such as pull-down assays or for ligating with streptavidin proteins. The PEG12 imparts the hydrophilicity of the compound. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	1-5ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	4ml
Binding yield of column	1mg

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid 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.

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at thelevel of PG by silica gel column purification

Kit Contents

Contents	IVD3182
Purification Times	50
Buffer ACL	250 ml
Buffer ACB*	300 ml
Buffer DCW1*	22 ml
Buffer DCW2*	10 ml
Proteinase K	540 mg
Protease Dissolve Buffer	30 ml
Carrier RNA	110 μg
Nuclease Free Water	20 ml
HiPure CFDNA Mini Columns	50
2 ml Collection Tubes	100
Extender Tube	50
Vac-Connector	50

Storage and Stability

Proteinase K, Carrier RNA 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.

Experiment Data

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,

Overview

• Ready-to-use
• Quantitative
• Highly Stable
• Precise
• Nineteen discrete fragments ranging from 100 bp to 10000 bp
• Higher intensity reference bands at 500 bp, 1000 bp and 5000 bp

The Norgen HighRangerPlus 100 bp DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our HighRangerPlus contains nineteen discrete fragments ranging from 100 bp to 10000 bp with higher intensity reference bands at 500 bp, 1000 bp and 5000 bp. This Ladder is ideal for size determination of digested DNA.

Contents:
1mL of premixed DNA ladder (0.5µg/10µL) in loading buffer (10mM EDTA, 10% glycerol, 0.015% bromophenol blue, and 0.17% SDS).

Details

Supporting Data

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HighRanger Plus 100 bp DNA Ladder (Cat# 12000) – 100 loads

Ladder Properties:

• Nineteen discrete fragments ranging from 100 bp to 10000 bp
• Higher intensity bands at 500 bp, 1000 bp and 5000 bp for easy reference

Fragment	Size (bp)	Mass (ng)
1	10000	42
2	8000	33
3	6000	25
4	5000	42
5	4000	35
6	3000	27
7	2500	22
8	2000	18
9	1500	13
10	1000	40
11	900	28
12	800	25
13	700	34
14	600	19
15	500	44
16	400	13
17	300	18
18	200	13
19	100	10

Recommended Use:
Mix thoroughly. For best results, load 10µL of DNA ladder per well. For precise mass determination with a densitometer, stain gel after electrophoresis using 0.5µg/mL ethidium bromide for 30-40 minutes. The table above shows the size and mass for each band based on 10µL ladder per well.

Storage: 
This kit is stable for 2 years after the date of shipment.

This ladder was standardized using 10µL of DNA per lane on a 0.8 cm thick, 13 x 15 cm, 1.0% agarose gel run in TAE buffer.