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.

Introduction

Usages:
Yeast extract is made from yeast, used in microbiological culture media as a source of nitrogen and B vitamins.

Technical specification:
Total nitrogen(TN)—————————≥10.0%
Amino nitrogen(AN) ————————≥4.0%
Moisture—————————————≤5.0%
Ash———————————————≤10.0%
Clˉ——————————————–＜2.0%


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

Specifications: 500g/bottle; 10kg/bag

500g/bottle; 10kg/bag

Introduction

Usages:

For the rapid detection and enumeration of coliform bacteria.

Principle:

Peptone and yeast extract powder provides carbon and nitrogen sources and trace elements; sodium chloride maintains osmotic equilibrium; agar as medium coagulant; dodecyl sulfate inhibit Gram-positive bacteria; chromogenic substrate and large intestine flora β- galactosidase-glucosidase specific reaction, hydrolysis of the substrate, the release of the color groups  produce green colonies on the light yellow plate.

Formulation (per liter):

Peptone :10g

Yeast extract powder: 3g

sodium chloride:5g

sodium lauryl sulfate:0.1g

Agar :12g

Chromogenic substrate 2.7g

Final pH 7.0 ± 0.2

How to use:

1. Weigh 32.8g of this product, adding 1L of distilled or deionized water , heated to boiling stirring until completely dissolved, dispensing into flask without autoclaving.

2,Take 25g or 25mL of sample with aseptic procedures, added to the flask containing 225mL of sterile phosphate buffered saline (or saline) is shaken thoroughly homogenized with a homogenizer or a 1:10 dilution 1min, then 1:10 dilution continue to select the appropriate serial dilutions of three, two plates each dilution was inoculated.

3, the use of pour method: The medium is cooled in a water bath to about 50 , sterilized petri dish having a diameter 90mm, 1ml samples were inoculated per dish, then poured into about 15ml of the above dissolution medium, and mix to solidify, upside down, 37 for 24 hours.

4, using surface inoculation: cooled to about 50 , shake devoted to the medium in sterile petri dish. Can be stored in the refrigerator for a day or stored at room temperature for several days (dark, 4 ). The sample was streaked method or filter method, 37 for 24 hours.

5, observe the results.

Quality control:

This product appear light yellow after pouring into plate, these strains were inoculated after 36 ± 1 18 ~ 24h culture growth in the following table.

Growth of bacteria were cultured bacteria numbers feature

Escherichia coli ATCC25922 good green colonies

Citrobacter ATCC8090 good green colonies

Salmonella typhimurium CMCC50115 good colorless colonies

Enterococcus faecalis ATCC29212 suppressed —–

Storage: Store at 10-30 , dark, cool and dry place, tighten the cap immediately after use. Storage period of two years.

1000mL