Introduction

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.

Details

Specifications

Features	Specifications
Main Functions	Co-isolation DNA and RNA(not include miRNA) from a single sample (cells, soft tissue, plant sample)
Applications	RT-PCR, cDNA synthesis, PCR andsecond-generation sequencing, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Soft tissue samples (viscera, excluding skin andmuscle), cultured cells and common plant tissues
Sample amount	Soft Tissue: < 30mg, Cell: <1 x 107, Plant: <100mg
Yield	DNA: 1 – 20 μg,  RNA: 3 – 100 μg

Principle

The Kits are designed to purify both genomic DNA and total RNA from the same cellor tissue sample. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column and bind DNA. Ethanol is added to the flow-through and the sample is applied to an RNA column. DNA/RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit. High-quality DNA is eluted in as little as 50µl water using the Kit.

Advantages

• High quality – high purity total RNA / DNA can be directly used in a variety of downstream applications
• Fast – column method can complete the extraction of several samples in 30 minutes
• Safe – no phenol chloroform extraction required
• Simultaneous extraction- simultaneously isolate DNA and RNA from one sample

Kit Contents

Contents	R511102	R511103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RLC	50 ml	200 ml
Buffer DW1	30 ml	150 ml
Buffer RW1	30 ml	150 ml
Buffer RW2*	20 ml	2 x 50  ml
RNase Free Water	10 ml	30 ml
Buffer AE	10 ml	50 ml

Storage and Stability

HiPure DNA/RNA Kit can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

This Kit is designed to purify genomic DNA and total RNA simultaneously from a single biological sample. Lysate is first passed through a DNA spin column to selectively isolate DNA and then through an RNA column to selectively isolate RNA. Pure DNA and RNA are purified from the entire sample, in contrast to other procedures where either the biological sample or the purified total nucleic acids is divided into two before being processed separately. The kit is compatible with small amounts of a wide range of animal cells and tissues.

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.

N-(endo-BCN-PEG2-amido-PEG3)-N-bis-(PEG3-Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is is a multi-functional PEG linker with six terminal propargyl groups and a BCN group. The propargyl groups enables formation of triazole linkage with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. The BCN group can react with azide -tagged compound or biomolecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

N-(endo-BCN-PEG2-amido-PEG3)-N-bis-(PEG3-Amino-Tri-(Propargyl-PEG2-ethoxymethyl)-methane) is is a multi-functional PEG linker with six terminal propargyl groups and a BCN group. The propargyl groups enables formation of triazole linkage with azide-bearing compounds or biomolecules in copper catalyzed Click Chemistry. The BCN group can react with azide -tagged compound or biomolecules. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.