Introduction

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Details

Specifications

Features	Specifications
Main FunctionsC	Co-isolation DNA and RNA from a single FFPE tissue sample
Applications	RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	No more than six 10µm sections of 150mm2 surface area or three 20µm sections of 150mm2 surface area.

Principle

FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. For RNA purification, transfer RNA Lysate to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer. For DNA purification, transfer DNA Lysate to an adsorption column and DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA was finally eluted with low-salt buffer.

Advantages

1.Use non-toxic dewaxing solution without contact with xylene
2.Obtain both DNA and RNA simultaneously from the same sample. Elute separately without affecting each other (Have the same steps and effects as top brand 80234, perfect substitute.)

Kit Contents

Contents	IVD5116
Purification Times	50 Preps
HiPure DNA Micro Column	50
HiPure RNA Mini Column I	50
2ml Collection Tubes	150
Proteinase K	50 mg
Protease Dissolve Buffer	5 ml
Buffer DPS	60 ml
Buffer FRL	15 ml
Buffer ATL	15 ml
Buffer RLC	15 ml
Buffer AL	15 ml
Buffer VHB	44 ml
Buffer RW2	25 ml
RNase Free Water	10 ml
Buffer AE	10 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.

Experiment Data

Description 

YesBlot™ Western Marker I is a ready-to-use mixture with ten IgG-binding proteins covering a wide range of molecular weights from 15 to 200 kDa in Tris-Glycine buffer. YesBlot™ Western Marker I performs dual functions. First, it contains 4 pre-stained proteins (10, 25, 45 and 70 kDa) for monitoring protein separation during SDS-PAGE, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the protein size. Second, ten IgG-binding proteins can be immuno-detected on film or by CCD imaging. YesBlot™ Western Marker I is compatible for chemiluminescent, fluorescent, chromogenic or other detection systems. In addition, YesBlot™ Western Marker I has two reference bands with enhanced intensity (at 30 kDa and 80 kDa). The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading. 

Features

• Direct visualization — 10 IgG-binding proteins for direct visualization on Western blots
• Pre-stained bands — 4 prestained proteins for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
• Wide range — 10 clear bands from 15 to 200 kDa for size estimation
• Quick reference — two enhanced bands (30 and 80 kDa)

Contents 

The YesBlot™ Western Marker I contains recombinant IgG binding proteins, prestained recombinant proteins, glycerol and SDS. 

Storage

4°C for 3 months
-20°C for 24 months

Overview

• Fast, reproducible and easy processing of samples
• High yield and high quality genomic DNA with no RNA or protein contamination
• DNA ready for any application including PCR, qPCR, genotyping and more

Norgen’s Cells and Tissue DNA Isolation Kits are designed for the rapid preparation of genomic DNA from cultured cells as well as various tissue samples and urine. The purified genomic DNA is fully digestible with all restriction enzymes tested, and is completely compatible with PCR and Southern Blot analysis.

Cells and Tissue DNA Isolation Kit (Spin Column)

Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The protocol can be completed in approximately 30 minutes for cells and within 90 minutes for tissues. Each kit contains sufficient materials for 50 preparations.

Cells and Tissue DNA Isolation Micro Kit (Micro)

Optimized for small inputs of cells and tissues, such as Laser-Captured Microdissection (LCM). Purification is based on spin column chromatography as the separation matrix. Norgen’s columns bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. Preparation time for a single sample is approximately 60 minutes, and each kit contains sufficient materials for 50 preparations.

Cells and Tissue DNA Isolation Kit (Magnetic Bead System)

Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. Preparation for 10 purifications is approximately 40 minutes of hands-on time.

Cells and Tissue DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)

Purification is based on the use of magnetic beads that bind DNA under optimized binding conditions. Norgen’s Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) allows for the isolation of genomic DNA from various types of animal tissues or cell samples. The Cells and Tissue DNA Isolation 96-Well Kit (Magnetic Bead System) also can be integrated with a robotic automation system.

Details

Supporting Data

Figure 1 / 9

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Kit Specifications
Maximum Input	20 mg of animal tissue Up to 150 μL of viral suspension 3 x 106 cells
Column Binding Capacity	> 50 μg
Average Yield	8 μg (from 1 x 106 HeLa Cells) 10 μg (from 10 mg kidney)
Elution Volume	50 – 200 μL
Analyte Purified	Genomic DNA, mitochondrial DNA, viral DNA
Format	Spin Column
Time to Complete 10 Purifications	30 min (cells) and 90 min (tissue)

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 1 year after the date of shipment when stored at room temperature.

Component	Cat. 53100 (50 preps)	Cat. 57300 (50 preps)	Cat. 59100 (50 preps)	Cat. 62500 (192 preps)
Lysis Buffer B	20 mL	20 mL	20 mL	1 x 40 mL 1 x 20 mL
Solution WN	18 mL	18 mL	18 mL	55 mL
Wash Solution A	18 mL	18 mL	–	–
Elution Buffer B	30 mL	8 mL	15 mL	1 x 30 mL 1 x 15 mL
Proteinase K	1.2 mL	1.2 mL	1.2 mL	–
Proteinase K in Sotrage Buffer	–	–	–	4 mL
Magnetic Bead Suspension	–	–	2 x 1.1 mL	8.5 mL
96-Well Plate	–	–	–	2
Spin Columns	50	–	–	–
Micro Spin Columns	–	50	–	–
Collection Tubes	50	50	–	–
Elution Tubes (1.7 mL)	50	50	50	–
96-Well Elution Plate	–	–	–	2
Adhesive Tape	–	–	–	2
Product Insert	1	1	1	1