Introduction

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from bacteria, yeast cells
Applications	RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Bacteria, yeast cells
Sample amount	Bacteria: <10^9；Yeast cells：<2 x10^7
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such as guanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein and other impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.

Advantages

• Fast – several samples can be extracted in 30 minutes
• High purity – the purified RNA can be directly used in various downstream applications
• High recovery – RNA can be recovered at the level of PG
• Good repeatability – silica gel column purification technology can obtain ideal results every time
• Broad spectrum – it can deal with various bacteria, including Gram-positive bacteria that are difficult to be lysed
• Sufficient components – the kit contains carried wall breaking enzymes and glass beads

Kit Contents

Contents	R418202	R418203
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
Glass Beads (0.1-0.6mm)	30 g	150 g
DNase I	600 μl	5 x 600 μl
DNase Buffer	6 ml	30 ml
Protease Dissolve Buffer	1.8 ml	15 ml
Buffer ATL	50 ml	200 ml
Buffer PHC	50 ml	200 ml
Buffer GXP2*	20 ml	100 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

Buffer PHC should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Buffer PHC up to 12 weeks) at room temperature(15–25°C) does not affect their performance. The remaining kit components can be stored at roomtemperature (15–25°C) and are stable for at least 18 months under these conditions.

Hipure Microbial RNA kit is suitable for extracting high-purity total RNA from bacterial/yeast culture medium. This kit combines high-efficiency Magzol Reagent (one-step extraction reagent) and silicagel column purification technology to complete the extraction of high-purity total RNA in only 40 minutes. The obtained RNA can be directly used in RT-PCR, Northern blot, poly-A + purification, nucleic acid protection and in vitro translation.

Overview

• Ensure optimal results for sensitive applications like NGS
• Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
• Purification and enrichment of intact urinary exosomes for functional studies.
• Bind and elute all RNA irrespective of size or GC content, without bias.
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• No phenol extractions, Proteinase K treatment, nor carrier RNA required.
• No time-consuming ultracentrifugation, filtration nor special syringes are required.
• No precipitation reagents nor overnight incubation required.
• Compatible with urine from any species.
• Pure exosomes are purified and are free-from any other RNA-binding proteins.
• Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary resin separation matrix.

Norgen’s EXTRAClean Urine Exosome Purification and RNA Isolation Mini Kit constitutes an all-in-one system for the purification of exosomes and the subsequent isolation of RNA from different urine sample volumes ranging from 250 μL to 1 mL. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The purified RNA is free from any protein-bound circulating RNA and of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, NGS application, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

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Minimum Urine Input	250 μL
Maximum Urine Input	1 mL
Size of Exosomes Purified	40 nm – 150 nm
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35-40 minutes
Average Yields	Variable depending on specimen

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

TCO-PEG8-DBCO is a heterobifunctional click chemistry reagent containing a TCO and a DBCO moiety. TCO group specifically and efficiently reacts with terrahydrazine at fast speed. DBCO is very reactive toward Azide through copper free click chemistry, the PEG spacer increases the aqueous solubility of the reagent. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

TCO-PEG8-DBCO is a heterobifunctional click chemistry reagent containing a TCO and a DBCO moiety. TCO group specifically and efficiently reacts with terrahydrazine at fast speed. DBCO is very reactive toward Azide through copper free click chemistry, the PEG spacer increases the aqueous solubility of the reagent. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.