Introduction

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA and miRNA from cell and tissue without MagZol reagent
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	Cell, Animal tissue, Plant tissue
Sample amount	Cells: ≤ 5 x 10^6, Animal tissue:<10mg
Elution volume	≥30μl
Time per run	≤40 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

Biological samples are first lysed and homogenized in a highly denaturing guanidine isothiocyanate-containing buffer, which immediately inactivates DNases and RNases to ensure isolation of intact DNA and RNA. The lysate is then passed through a Mini spin column. This column, in combination with the high-salt buffer, allows selective and efficient binding of genomic DNA. Flow-through from the column is digested by Proteinase K in the presence of ethanol. This optimized digestion, together with the subsequent addition of further ethanol, allows appropriate binding of total RNA, including miRNA, to the column. Contaminants are efficiently washed away and high-quality RNA is eluted.

Advantages

• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – several samples can be extracted in 40 minutes
• High applicability – samples including animals, plants, bacteria, cells, etc.
• High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
• Safe – no phenol chloroform extraction and no use of Trizol reagent

Kit Contents

Contents	R431102	R431103
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
Proteinase K	48 mg	240 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer RLC	40 ml	200 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	60 ml

Storage and Stability

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

The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples.

OverView

The PCR Decontamination Kit can remove contaminating DNA in PCR master mixes, without reduction of PCR sensitivity.

• The double-strand specific property of the dsDNase allows decontamination with primers and probe present.
• Efficient for end-point PCR, probe-based qPCR, and some SYBR based qPCR mixes.
• Contaminating bacterial DNA can be reduced to levels below the detection limit.
• Fast and easy protocol.
• Flat NTCs (No Template Controls).

Decontamination of master mixes without reduction of sensitivity has always been a challenge. Especially when minor amounts of DNA are targeted, contaminating DNA is a major problem. Any loss of sensitivity in the qPCR assay caused by the decontamination protocol is unacceptable.

In Figure 1, it is demonstrated that the PCR decontamination kit can remove contaminating DNA from a qPCR mix to non-detectable levels (flat NTC), without affecting the sensitivity of the qPCR.

Kit Contents

• DTT (Inactivation Aid)
• dsDNase

The PCR Decontamination Kit can remove contaminating DNA in PCR master mixes, without reduction of PCR sensitivity.

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.

Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings