【IT1100】EzRNA™ T7 High Yield RNA Synthesis Kit (Ψ-UTP), 50 RXN

Overview

• Isolate high quality, high yield plasmid DNA
• Plasmid DNA is ready for various downstream applications including restriction digestion, bacterial transformation, sequencing and more
• Available in 4 formats: MiniPrep, MiniPrep (Magnetic Bead System), 96-Well MiniPrep (Magnetic Bead System), and MaxiPrep

These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.

Plasmid MiniPrep Kit

This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.

Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)

Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.

Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.

Plasmid MaxiPrep Kit

This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	1.5 mg
Average Yield from 100 mL Culture*	0.4 – 1.0 mg
Final Endotoxin Levels	< 0.1 EU/µg DNA
Time to Complete 4 Purifications	1.5 hours
Size of Plasmids Purified	Up to 13,000 bp

 * Depends on high-copy DNA and low copy DNA

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. The RNase vial should be stored at -20°C upon arrival. Once RNase has been added to the Resuspension Solution AZ the solution should be stored at 4°C. This kit is stable for 1 year after its date of shipment.

Component	Cat. 13300 (50 preps)	Cat. 46400 (250 preps)	Cat. 46500 (4 preps)	Cat. 46600 (20 preps)	Cat. 60300 (50 preps)	Cat. 63000 (192 preps)
Resuspension Solution AZ	12 mL	60 mL	20 mL	100 mL	12 mL	2 x 20 mL
Lysis Buffer N	40 mL	80 mL	40 mL	2 x 80 mL	40 mL	2 x 40 mL
Buffer TN	20 mL	130 mL	55 mL	2 x 130 mL	20 mL	1 x 55 mL 1 x 20 mL
Wash Solution E	12 mL	2 x 18 mL	–	–	–	–
Elution Buffer K	8 mL	30 mL	–	–	8 mL	2 x 8 mL
Wash Solution J	–	–	25 mL	3 x 25 mL	–	–
Elution Buffer J	–	–	24 mL	120 mL	–	–
RNase A	1 vial	1 vial	1 vial	1 vial	1 vial	1 vial
Magnetic Bead Suspension	–	–	–	–	1 x 1.1 mL	4 x 1.1 mL
Spin Columns	50	250	–	–	–	–
Collection Tubes	50	250	–	–	–	–
DNA Maxi Spin Columns with Collection Tubes (Clear ring in column)	–	–	4	20	–	–
Maxi Spin Filter Columns with Collection Tubes (Grey ring in column)	–	–	4	20	–	–
96-Well Plate	–	–	–	–	–	2
Elution Tubes (1.7 mL)	50	250	–	–	50	–
Elution Tubes (50 mL)	–	–	4	20
96-Well Elution Plate	–	–	–	–	–	2
Adhesive Tape	–	–	–	–	–	2
Product Insert	1	1	1	1	1	1

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile urine input ranges
• No phenol extractions
• No carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Concentrate circulating RNA and exosomal RNA into flexible elution volumes
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating RNA, including exosomal RNA as well as viral RNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Recent evidence indicates that cell-free circulating RNA (cf-RNA) including exosomal RNA in urine contains valuable information for the discovery of biomarkers that can help for the early detection of certain cancer types and for monitoring the disease status as well as for the detection of any infectious pathogens. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acid for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum but with a lower concentration; (3) Nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).

Urine Cell-Free Circulating RNA Purification Mini Kit

For sample volumes ranging from 250 µL to 2 mL.

Urine Cell-Free Circulating RNA Purification Midi Kit

For sample volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.

Urine Cell-Free Circulating RNA Purification Maxi Kit

For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	10 mL
Maximum Urine Input	30 mL
Size of RNA Purified	All sizes including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	40-45 minutes
Average Yields	Variable depending on specimen

*Please check page 6 of the product insert for average yields and common RNA quantification methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are 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.

Component	Cat. 56900 (50 preps)	Cat. 57000 (20 preps)	Cat. 57100 (10 preps)
Binding Solution K	25 mL	75 mL	1 x 75 mL 1 x 25 mL
Lysis Buffer A	30 mL	20 mL	20 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Spin Columns	50	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1