Overview

• Can be used for the clean up of both RNA and DNA from enzymatic reactions, labeling etc.
• Purifies all sizes of RNA, from large mRNA down to microRNA (miRNA)
• Purifies all sizes of DNA, from small PCR products to plasmids to genomic DNA
• Removes endotoxins for transfection of injection ready RNA or DNA
• Rapid and efficient spin-column format (20 minutes)
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s CleanAll DNA/RNA Clean-Up and Concentration Micro Kit provides a rapid method for the purification, cleanup and concentration of RNA or DNA from different isolation methods or upstream applications.   The kit can be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions. 

The CleanAll Kit purifies RNA from phenol/guanidine-based protocols or from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. Furthermore, endotoxins can be removed from previously purified RNA solutions.

The kit can be used to clean up DNA from digestions, ligations, PCR reactions, labeling reactions, DNA modification reactions and staining. The kit also provides a protocol for the rapid removal of endotoxins from previously purified DNA down to 0.1 EU/µg DNA or less.

Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).

Purify all sizes of DNA, from small PCR products, native or linearized plasmids, to genomic DNA. Using an alternative protocol, even oligonucleotides and smaller DNA fragments can be purified.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	50 μg RNA 10 μg DNA
Maximum Column Loading Volume	600 μL
Minimum Elution Volume	20 μL
Time to Complete 10 Purifications	20 minutes
Size of RNA Purified	All sizes, including small RNA (<200)
Size of DNA Purified	≥ 100 bp*
Average Recovery	≥90% for RNA ≥90% for DNA 100 bp to 10 kbp ≥75% for DNA ≥ 10 kbp

* Applicable to smaller fragments or oligonucleotides with alternative protocol

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years from the date of shipment.

Component	Cat. 23800 (50 preps)
Binding Buffer H	60 mL
Wash Solution K	27 mL
Elution Buffer L	15 mL
Micro Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Description

Specifications

Clone	IHC269
Source	Mouse Monoclonal
Positive Control	Spleen
Dilution Range	1:200

TIGIT is an immune receptor present on some T cells and Natural Killer cells. TIGIT binds with high affinity to the poliovirus receptor (PVR) which causes increased secretion of IL10 and decreased secretion of IL12B and suppresses T cell activation by promoting the generation of mature immunoregulatory dendritic cells. Through the CD226/TIGIT-PVR pathway, TIGIT regulates T cell mediated immunity. In cancer, TIGIT and PD-1 have been shown to be over-expressed on tumor antigen-specific CD8+ T cells and CD8+ tumor infiltrating lymphocytes (TILs) from individuals with melanoma. Blockade of TIGIT and PD-1 led to increased cell proliferation, cytokine production, and degranulation of tumour antigen-specific CD8+ T cells and TIL CD8+ T cells. It can be considered an immune checkpoint.

Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular nucleic acid fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp(Nucleic Acid). The HiPure Circulating Nucleic acid Micro Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be either fresh or frozen (provided that they have not been frozen and thawed more than once). Free-circulating cell-free DNA, RNA or viral nucleic acids are eluted in Nuclease Free Water, ready for use in amplification reactions or storage at -30 to -15°C. Purified nucleic acids are free of proteins, nucleases, and other impurities.

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 0.6ml plasma,serum, body fluids
Applications	qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Serum, plasma and other cell-free fluid samples
Sample amount	0.6ml
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based onsilica column purification. The sample is lysed and digested with lysate andprotease, DNA is released into the lysate. Transfer to an adsorption plate andfilter column. Nucleic acid is adsorbed on the membrane, while protein is notadsorbed and is removed with filtration. After washing proteins and otherimpurities, 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 recoveredat the level of PG by silica gel column purification

Kit Contents

Contents	D318002	D318003
Purification Times	50 Preps	250 Preps
Buffer ACL	40 ml	200 ml
Buffer DCW1	22 ml	110 ml
Buffer DCW2*	20 ml	2 x 50 ml
Proteinase K	34 mg	180 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Carrier RNA	110 μg	310 μg
Nuclease Free Water	10 ml	30 ml
HiPure CFDNA Mini Columns	50	250
2 ml Collection Tubes	100	5 x 100

Storage and Stability

Proteinase K, Carrier RNAshould be stored at 2-8°C upon arrival. However, short-term storage (up to 12weeks) at room temperature (15-25°C) does not affect their performance. Theremaining kit components can be stored dry at room temperature (15-25°C) andare stable for at least 18 months under these conditions. The entire kit can bestored 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.

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