Extraction Method

• RNAsafeguard Reagent MORE
  Product Details：

  Component	Amount
  RNAsafeguard Reagent	120 ml
  Manual	1

      RNAsafeguard is an aqueous, non-toxic tissue storage reagent that rapidly permeates tissue to stabilize and protect cellular RNA in situ in unfrozen specimens. Tissue pieces are harvested and immediately submerged in RNAsafeguard for storage without jeopardizing the basic structure of tissue cell and the quality or quantity of RNA. RNAsafeguard eliminates the need to immediately process tissue specimens or to freeze samples in liquid nitrogen or dry ice for later processing.

      RNAsafeguard preserves RNA in tissues for up to 1 day at 37 °C, 1 week at 25 °C, and 1 month or more at 4 °C. Tissues can also be stored at –20 °C or at –80 °C long-term.

      RNAsafeguard has been extensively tested on tissues from several vertebrate species, including brain, heart, kidney, liver, skeletal muscle,etc, at the same time tissue with all kinds of pathological damage can do as well. RNAsafeguard is also effective for tissue culture cells and white blood cells. RNAsafeguard is compatible with most RNA isolation methods, including TRI Reagent.

  Product Features：

  Storage and Stability

      Store RNAsafeguard at room temperature for at least 2 years. If a precipitate develops in the RNAsafeguard, warm the solution to 37 °C and agitate to redissolve it.

  Storage of Samples in RNAsafeguard

  1. Storage at –80 °C is recommended for long-term storage. Incubate samples at 4 °C overnight, and then remove them from RNAsafeguard before storage at –80 °C to prevent the formation of salt crystals. Samples can subsequently be thawed at room temperature and refrozen without affecting the amount or the integrity of the recoverable RNA.

  2. Storage at –20 °C is recommended for long-term storage. Incubate samples at 4 °C overnight, then transfer to – 20 °C.Crystals may form in the storage buffer; which will not affect subsequent RNA isolation. If crystals are a concern, remove the RNAsafeguard prior to storing the samples at –20 °C. Samples can subsequently be thawed at room temperature and refrozen without affecting the amount or the integrity of the recoverable RNA.

  3. Storage at 4 °C is recommended for up to 1 month without any experimental evidence of RNA degradation.

  4.  Storage at ambient temperature RNA isolated from samples stored at 25 °C for one week is intact. RNA from samples stored at 25 °C for two weeks appears slightly degraded, but still of sufficient quality for nuclease protection assay or RT-PCR analysis.

  5. RNA isolated from samples stored at 37 °C is intact after 24-hour incubation, but is partially degraded after a 3 day incubation.

• pBIOZOL Plant Total RNA Extraction Reagent MORE
  Product Details：

  Kit Components

  Cat#	BSC55S1	BSC55M1
  pBIOZOL	30ml	100ml
  Handbook	1copy	1copy

  Introduction 

      pBIOZOL provides a simple, rapid and effective method for isolation of high quality total RNA from plant tissues, especially those rich in polyphenolics or starch. Use of pBIOZOL results in high yields of high quality total RNA from plant tissues.

   

  Product Features：

  Storage and transportation

  •  The reagent should be stored dry at 4 ℃; the reagent can be stored for up to 6 months from date of delivery, if stored properly.
  •  The reagent can be transported at room temperature.

  Attention

      pBIOZOL contains β-mercaptoethanol and sodium azide. Use the reagent in a fume hood. Wear gloves and eye protection when handling the reagent and solutions containing the reagent.

  Technical Information

  Sample size	Reaction volume
  ≤100mg	0.5ml
  0.1 to 5g	5ml per 1g sample

  Working site

  fume hood

  RNase-free preparation

  • Always wear disposable gloves and change them frequently.
    • Use good microbiological technique to avoid contamination.
    • Use sterile, disposable plastic ware; soak in 0.1% DEPC overnight and autoclave.
    • Use pipettes specially reserved for RNA work.
  • Use aerosol-resistant pipette tips to reduce sample-to-sample contamination or reagent contamination.
    • Bake glassware at 150℃ for 4 hours.
  • Soak non-disposable plasticware in 0.5 M NaOH for 10 minutes, rinse thoroughly with RNase-free water.
    • Prepare RNase-free water by drawing the water into RNase-free containers.
    • Adding diethylpyrocarbonate (DEPC) to a final concentration of 0.01% (v/v). Let stand overnight and autoclave. Use this water to prepare RNase-free solutions.

  • All lab ware or solution related to the experiment should be RNase-free.
• dBIOZOL Genomic DNA Extraction Reagent MORE
  Product Details：

      dBIOZOL is a complete and ready to use reagent for the isolation of genomic DNA from samples of animal and plant origin. The dBIOZOL procedure is based on the use of a novel chaotropic-detergent lysing solution that hydrolyzes RNA and allows the selective precipitation of DNA from a cell lysate. The advanced DNA isolation method combines both reliability and efficiency with simplicity of the isolation protocol, which is fast and permits isolation of genomic DNA from a large number of samples of small or large volumes In general, 0.5-4 μg genomic DNA per mg tissue can be acquired from up to 80 mg tissue with dBIOZOL

      The isolated DNA can be used, without additional purification, for PCR/Real time PCR， sequencing，Southern blot， mutant analysis，SNP and other molecular biology and biotechnology applications.

  Principle

      During the isolation, a biological sample is lysed (or homogenized) in dBIOZOL and the genomic DNA is precipitated from the lysate with isopropanol. Following an ethanol wash, DNA is solubilized in elution buffer. The procedure can be completed in 40 minutes.

  Product Features：

      STABILITY: dBIOZOL is stable at room temperature for at least two years after the date of purchase. HANDLING PRECAUTIONS: dBIOZOL contains irritants. Handle with care, avoid contact with skin, and eye .In case of contact: Wash skin with a copious amount of water and seek medical attention.

  PROTOCOL

  The protocol includes the following steps:

  1.        LYSIS\HOMOGENIZATION: 0.6ml dBIOZOL+20 ～ 80 mg tissue(animal or plant), 105-108 cells(culture cells or white cells in whole blood)

  2.        CENTRIFUGATION

  Sediment the homogenate for 10 minutes at 13,000 g at 4-25 C. Following centrifugation, transfer the resulting viscous supernatant to a fresh tube.

  3.        DNA PRECIPITATION: lysate + 0.7ml isopropanol.

  4.        DNA WASH:1 ml 75% ethanol

  5.        DNA SOLUBILIZATION: 8 mM NaOH or water.

  Experimental data:

  ⊕Absorbance analysis

      Get some DNA，diluted in a advisable factor with elution buffer. Survey the OD260   ，OD280 and OD320.

  Expressions：concentration（μg/ml）＝50×OD260×dilution fact Target： 2.0≥OD260-320/ OD280-320≥1.7

  Notice: 1.0≥OD260≥0.1, the result of ratio is much reliable.

  ⊕Agarose Gel Analysis 0.8～1％ Agarose gel

  

  The fragment of the photography with 1,2,3,4 is rat kidney brain liver and heart,respectively,and with5,6 is plant leaf and soybean.

  Manual download
• BIOZOL Total RNA Extraction Reagent MORE
  Product Details：

  Cat#	BSC51S1	BSC51M1
  BIOZOL-total RNA Extraction reagent	30 ml	100 ml
  Handbook	1copy	1copy

  General information

      BIOZOL Reagent is a ready-to-use reagent for the isolation of total RNA from animal or plant tissue or from bacterium. The operation of the kit is very simple and easy. First, add the sample into BIOZOL for complete lyses. Second, add chloroform into the solution and then centrifuge the sample tube. Then the homogenate will separate into form three phases: upper aqueous phase, interphase phase and organic phase. RNA remains exclusively in the aqueous phase; the RNA is recovered by precipitation with isopropyl alcohol.

      Since the operation of BIOZOL is very easy, it can realize the extraction for several samples simultaneously. And total RNA isolated by BIOZOL Reagent is free of protein and DNA contamination. It can be used for Northern blot analysis, dot blot hybridization, poly (A)+ selection, in vitro translation, RNase protection assay, RT-PCR assay, cDNA library building and other RNA research.

  RNA Extraction Procedure

   

  
  

  Product Features：

  Storage and Transportation

  ◆   Recommended to store at 2-8℃ and the stability is one year. Recommended to avoid light source so as to achieve optimized effect. Stored at room temperature for a while will not influence the effect.

  ◆   It can be shipped at room temperature which does not affect its stability.

  Important Notes

      RNase can be introduced accidentally into the RNA preparation at any point in the isolation procedure through improper technique. Because RNase activity is difficult to inhibit, it is essential to prevent its contamination. The following guidelines should be observed when working with RNA.

  ◆    Always wear gloves and respirator in order to prevent RNA degradation.

  ◆    Use sterile, disposable plastic ware and automatic pipettes.

  ◆    All plastic wares and tips should be RNase-free as recommended done as below: After washed, metal ware items can be baked at 150°C for 6－8 hours, while  tips or glass and plastic items can be soaked for 2 hours at 37 °C or whole night (12hrs) at room temp in 0.1%(v/v) diethylpyrocarbonate (DEPC) solution, and autoclaved for 20mins in 151bf/in2 (1.034x105 Pa) autoclave and then baked at 60°C.

  ◆    Build a separate RNA laboratory with the only instruments if possible.

  Experimental data:

   

  Total RNA pictures above are strictly operated according to Biozol extraction protocol.

  
  

  Manual download