Propargyl-PEG6-NHS ester

Introduction

029080 Glycerin

Usage:Additive Reagent for Culture Media.

Specification:1ml*10vials

1ml*10vials

OverView

HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human  total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results  indicate that HL-dsDNase has minimal impact on RNA quality and quantity.

HL-dsDNase is an engineered version of dsDNase that is rapidly and completely inactivated by incubation for 5 minutes at 58°C with 1 mM DTT at pH 8.0 or above. Chemical inactivation in downstream compatible RT or PCR buffers allows milder heat inactivation and, in some cases, skipping heat inactivation altogether. This makes HL-dsDNase very useful for removal of DNA from RNA preparations since the enzyme may be inactivated using various strategies with reduced risk of auto-degradation of  RNA in the presence of magnesium, making HL-dsDNase an ideal enzyme when working with small volumens of RNA.

HL-dsDNase is also an excellent choice for removal of unwanted external DNA from samples prior to analysis of nucleic acids protected by biological membranes, e.g., bacteria, viruses, and sperm. Especially if using downstream analysis methods that might be affected by host cell DNA, as metagenomic sequencing and STR-profiling. The easy inactivation of HL-dsDNase makes it a fast and efficient alternative to methods as differential extraction, where sample material is often lost.

Recommended applications:

• Removal of genomic DNA from RNA preparations
• Improved removal of host-cell DNA from sperm cells prior to STR-profiling

Key advantages with HL-dsDNase:

• Double-strand DNA specific endonuclease
• Easily heat-inactivated by very moderate heat treatment
• High specific activity
• Useful for removal of DNA from RNA preps

Properties

HL-dsDNase is especially developed to remove contaminating genomic DNA from RNA preparations. Figure 1 shows that HL-dsDNase can reduce 50 ng of human gDNA to levels non-detectable by qPCR. In figure 2, a human  total RNA sample was treated with HL-dsDNase and analysed on the Bio-Rad Experion™ System. The results  indicate that HL-dsDNase has minimal impact on RNA quality and quantity.

Overview

• Sequentially isolate and purify total RNA and DNA from a single sample
• Two column system: one for DNA and one for RNA
• The RNA column is for the purification of total RNA including microRNA
• No need to split the lysate, or to use phenol or precipitation methods
• Rapid and efficient spin column procedure – it takes only 30 minutes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a rapid method for the isolation and purification of total RNA and DNA sequentially from a single sample of cultured animal cells and tissues, blood, bacteria, yeast, or fungi. The lysate is passed over two columns: 1) a DNA column and 2) an RNA column. Total RNA of all sizes is purified, including microRNA. Both DNA and RNA are of the highest purity and yield.

These kits are ideal for researchers who are interested in studying the genome and transcriptome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/DNA Purification Kits are especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as they eliminate the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and DNA are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications

RNA/DNA Purification Kit (Spin Column)

Maximum column binding capacity of 50 μg for RNA and 20 μg for DNA.

RNA/DNA Purification Micro Kit (Micro)

The purified RNA and DNA fractions can be eluted in as little as 20 μL. Ideal for cell number inputs of 500,000 and as little as 5 cells. Maximum column binding capacity of 35 μg for RNA and 10 μg for DNA.

Details

Supporting Data

Figure 1 / 4

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Kit Specifications
Maximum Column Binding Capacity	50 μg for RNA 20 μg for DNA
Maximum Column Loading Volume	650 μL
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Maximum Amount of Starting Material: Animal Cells Animal Tissues Blood Bacteria Yeast Fungi Plant Tissues	5 x 106 cells 25 mg (for most tissues)** 200 μL 1 x 109 cells 1 x 108 cells 50 mg 50 mg
Time to Complete 10 Purifications	30 minutes
Average Yield*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg)	10-15 μg RNA 2-4 μg DNA 30-35 μg RNA 4-6 μg DNA

*Average Yield will vary depending upon a number of factors including species, growth conditions used, and development stage.

**Tissue inputs of up to 40 mg may be used, however for inputs greater than the recommended 25 mg, cross-contamination of the RNA and DNA fractions is possible.

Storage Conditions and Product Stability
Store Proteinase K at -20°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 48700 (50 preps)	Cat. 50300 (50 preps)
Buffer SKP	40 mL	40 mL
Wash Solution A	2 x 38 mL 1 x 18 mL	2 x 38 mL 1 x 18 mL
Elution Solution A	6 mL	6 mL
Elution Buffer F	15 mL	6 mL
RNase-Free Water	40 mL	40 mL
Proteinase K	2 x 12 mg	2 x 12 mg
gDNA Purification Columns	50	–
gDNA Purification Micro Columns	–	50
RNA Purification Columns	50	–
RNA Purification Micro Columns	–	50
Collection Tubes	100	100
Elution Tubes (1.7 mL)	100	100
Product Insert	1	1