tert-Butyl (2-(prop-2-yn-1-yloxy)ethyl)carbamate

Overview

• Extract high quality & quantity total RNA including miRNA
• No phenol step required – isolate all RNA in one fraction
• Rapid processing in under 40 minutes
• Isolate total RNA from a wide variety of specimens
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s Total RNA Purification Maxi Kit provides a rapid method for the isolation and purification of total RNA from cultured animal cells, tissue samples, blood, bacteria, yeast, fungi, plants, and viruses. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Norgen’s Purification Technology

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. The RNA is preferentially purified from other cellular components such as proteins without the use of phenol or chloroform. The process involves first lysing the cells or tissue of interest with the provided Buffer RL (please see the flow chart on page 4). Ethanol is then added to the lysate, and the solution is loaded onto a maxi spin column. Norgen’s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the column, while the contaminating proteins will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed with the provided Wash Solution A in order to remove any remaining impurities, and the purified total RNA is eluted with the Elution Solution A. The purified RNA is of the highest integrity, and can be used in a number of downstream applications.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	1.5 mg
Maximum Column Loading Volume	20 mL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material
Liver Tissue	100 mg
Heart Tissue	50 mg
Kidney Tissue	100 mg
Brain Tissue	250 mg
Lung Tissue	100 mg
Whole Blood	2 – 10 mL*
Bacteria	2.5 x 10 10 cells
Yeast	2 x 108 cells
Fungi	1 g
Plant Tissues	1 g
Time to Complete 4 Purifications	40 minutes
Average Yield: HeLa Cells (5 x 107 cells) E. coli (2.5 x 1010 cells)	~750 μg ~1.5 mg

*For isolating total RNA from purified leukocytes

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

Component	Cat. 26800 (8 preps)
Buffer RL	2 x 40 mL
Wash Solution A	4 x 38 mL
Elution Solution A	3 x 20 mL
RNA Maxi Spin Columns (with collection tubes)	8
Elution Tubes (50 mL)	8
Product Insert	1

Overview

• Ready-to-use
• Quantitative
• Highly Stable
• Precise
• Ten discrete fragments ranging from 300 bp to 5000 bp

The Norgen MidRanger 1 kb DNA Ladder is prepared to ensure quality and batch-to-batch consistency. Our MidRanger contains ten discrete fragments ranging from 300 bp to 5000 bp. This Ladder is ideal for sizing larger PCR products (>1kb) and most cloning applications.

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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MidRanger 1kb DNA Ladder (Cat# 11700) – 100 loads

Ladder Properties:
• Ten discrete fragments ranging from 300 bp to 5000 bp

Fragment	Size (bp)	Mass (ng)
1	5000	88
2	4000	78
3	3000	67
4	2500	58
5	2000	50
6	1500	43
7	1000	25
8	700	34
9	500	33
10	300	24

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: 

Stable at room temperature. For longer term storage, -20°C is recommended.

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.

DBCO-PEG4-Butyraldehyde is a click chemistry PEG reagent containing a terminal butylraldehyde group. Butyraldehyde reacts with hydrazide to form a stable hydrazone linkage or with alkoxyamines to form a stable oxime bond. The hydrophilic PEG spacer arm improves water solubility and provides a long and flexible connection that minimizes steric hindrance involved with ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research use only.

DBCO-PEG4-Butyraldehyde is a click chemistry PEG reagent containing a terminal butylraldehyde group. Butyraldehyde reacts with hydrazide to form a stable hydrazone linkage or with alkoxyamines to form a stable oxime bond. The hydrophilic PEG spacer arm improves water solubility and provides a long and flexible connection that minimizes steric hindrance involved with ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research use only.