Introduction

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from FFPE tissue and section samples (with DNase)
Applications	RT-PCR, quantitative RT-PCR, Northern hybridization, Poly A purification, nucleic acid protection and in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology, DNase
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE tissue sample
Sample amount	6mg
Yield	20μg
Elution volume	≥10μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer 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.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 60 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Efficient DNA removal – unique method to effectively remove genomic DNA

Kit Contents

Contents	R414402	D414403
Purification Times	50 Preps	250 Preps
HiPure RNA Micro Columns	50	250
2ml Collection Tubes	50	250
Buffer DPS	60 ml	250 ml
Buffer FRL	15 ml	60 ml
Buffer RLC	15 ml	60 ml
Buffer RWC*	10 ml	50 ml
Buffer RW2*	20 ml	2 x 50 ml
DNase I	600 µl	5 x 600 µl
DNase Booster Buffer	1.5 ml	6 ml
Protease Dissolve Buffer	1.8 ml	10 ml
Proteinase K	24 mg	120 mg
RNase Free Water	10 ml	20 ml

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Overview

• Rapid and simple procedure
• Excellent quality and yield of DNA
• Process a broad spectrum of plant species and filamentous fungi
• Isolate total DNA including pathogen DNA without phenol 
• Available in spin column format and 96-well format for high throughput applications

These kits provide a rapid method for the isolation and purification of total DNA from a wide range of plant and fungal species. Total DNA, including genomic DNA, mitochondrial DNA and chloroplast DNA can be purified from fresh or frozen plant tissues, plant cells or fungi samples using this kit. Purified DNA samples can be used for the detection of viral pathogens, as viral DNA is isolated with the plant/fungi DNA. The purified DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, SNP, Southern blotting and sequencing.

Plant/Fungi DNA Isolation Kit (Spin Column)

Complete 10 purifications in 45 minutes. This kit offers a maximum loading volume of 650 μL per column, and a maximum binding capacity of 50 μg per column.

Plant/Fungi DNA Isolation 96-Well Kit (High Throughput)

For high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system. Complete 96 purifications in 50 minutes. This kit offers a maximum loading volume of 500 μL per well, and a maximum binding capacity of 50 μg per well.

Plant/Fungi DNA Isolation Kit (Magnetic Bead System)

The DNA is bound to the surface of the magnetic beads under optimized buffer conditions and released using a low salt buffer system. The Plant DNA Isolation Kit (Magnetic Bead System) can be easily adapted to automated magnetic bead separation instruments and work stations.

Plant/Fungi DNA Isolation 96-Well Kit (High Throughput Magnetic Bead System)

For high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.

Details

Supporting Data

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Kit Specifications – Spin Column
Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Maximum Amount of Starting Material:Plant TissuesFungi (wet weight)	100 mg 100 mg
Average Yields* 50 mg Tomato Leaves 50 mg Grape Leaves 50 mg Peach Leaves 50 mg Plum Leaves 50 mg Pine Needles Botrytis cinerea (50 mg wet weight) Fusarium sp. (50 mg wet weight) Aspergillus niger (50 mg wet weight)	18 µg 10 µg 10 µg 10 µg 5 µg 1.5 µg 2 µg 4 µg
Time to Complete 10 Purifications	45 minutes

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

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

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature, except for the RNAse which should be stored at -20°C. This kit is stable for 1 year after the date of shipment.

Select Plants and Fungi that can be used with the Plant/Fungi DNA Purification Kits

Plants	Plants (Cont’d)	Fungi
Tomato	Turnip	Aspergillus niger
Grape	Chinese Cabbage	Mucor racemosus
Peach	Radish	Cladosporium cladosporioides
Plum	Komatsuna	Fusarium oxysporum
Pine Needles	Apricot	Penicillium sp.
Raspberry	Sweet Potato	Botrytis cinerea (Botryotinia fuckeliana)
Strawberry	Hydrangea	Pichia sp.
Legumes	Fig	Rhizopus oryzae
Prosopis cineraria (Ghaf)	Turf	Alternaria tenuissima
Sorghum grass	Cherry	Fusarium sp.
Tobacco	Saintpaulia
Arabidopsis	Lotus
Lichen	Carrot
Corn seed	Hansen
Sunflower seed	Pistachio
Olive seed
Soybean seed

Component	Cat. 26200 (50 preps)	Cat. 26250 (250 preps)	Cat. 26900 (192 preps)	Cat. 58200 (50 preps)	Cat. 62400 (192 preps)
Lysis Buffer L	30 mL	1 x 30 mL 2 x 60 mL	2 x 60 mL	60 mL	2 x 60 mL
Binding Buffer I	7 mL	1 x 7 mL 1 x 25 mL	25 mL	7 mL	25 mL
Solution WN	18 mL	1 x 18 mL 1 x 55 mL	55 mL	18 mL	55 mL
Wash Solution A	38 mL	2 x 38 mL	2 x 38 mL	–	–
Elution Buffer B	15 mL	30 mL	30 mL	8 mL	30 mL
RNAse A	1 vial (80 μL)	5 vials (5 x 80 μL)	1 vial	1 vial	1 vial
Magnetic Bead Suspension	–	–	–	4 x 1.1 mL	2 x 8.5 mL
Filter Columns	50	250	–	–	–
Spin Columns	50	250	–	–	–
Collection Tubes	100	500	–	–	–
96-Well Plate	–	–	2	–	2
96-Well Collection Plate	–	–	2	–	–
Adhesive Tape	–	–	4	–	2
Elution Tubes (1.7 mL)	50	250	–	50	–
96-Well Elution Plate	–	–	2	–	2
Product Insert	1	1	1	1	1

Introduction

Usages:

For antimicrobial susceptibility testing.

Principle:

Beef extract powder and acid hydrolyzed casein protein provides nitrogen, vitamins and amino acids; soluble starch absorption of toxic metabolites; agar as medium coagulant.

Formulation(per liter):

Agar 17g

Beef extract 2g

Soluble starch 1.5g

Acid hydrolysis of casein 17.5g

Final pH7.3 ± 0.2

How to use:

1.Suspend 38g in 1 L of distilled water , stirring heated to boiling until completely dissolved, dispensing flask, 121 autoclave for 15min.

2.Diluted and treated samples.

Storage: Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.

500g