RNall Kit

 22,959,600 41,163,600

RNall kit Provides two time-saving, reliable and meticulous methods for RNA and total RNA (including miRNA) isolation from various sample types including animal tissues (lipid and fibrouse), animal cells, PBMC and bacteria. However, we suggest using RNJia Bacteria Kit, RNJia Lipid Kit and RNJia Fibrous kits, which are specialized for RNA isolation from bacteria, lipid and fibrous tissues to reach optimum results, while RNall is capable of total RNA isolation (including miRNA) from these sample types if suitable enzymes are purchased. RNall kit is based on spin column technology for isolation of concentrated, highly purified and intact total RNA (including miRNA), which is suitable to use for variety of downstream processes such as Northern blot experiments, Real-time PCR, RNA Sequencing, Micro array and etc. In the kit process, two methods are possible: phenol- chloroform based (RNSol) and phenol-chloroform free based (RLB). However, it is recommended to use RLB for situations in which the purified RNA will be used in sensitive downstream applications.

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Description

Kit content

Component 50 preps 100 preps
RNSol Reagent 50 ml 100 ml
RLB 50 ml 100 ml
TWB1 (concentrate) 17 ml 35 ml
TWB2 (concentrate) 15 ml 27.5 ml
Nuclease-free Water 5 ml 10 ml
Spin Column 50 100
Collection Tube 50 100

Storage

Shipment condition is checked by ROJETechnologies. After arrival, RNSol Reagent should be kept dry, at 2-8 °C and all remaining reagent should be kept at room temperature (15-25 °C). When storage condition is as directed, all reagents are stable until expiration date, as indicated on the kit box.

Intended Use

The RNall Kit provides the components and procedures necessary for purifying RNA and miRNA from cultured cells, animal tissue, PBMC and bacteria cells. Notice that, RNall is intended for molecular biology applications not for diagnostic use.  We recommend all users to study RNA experiments guideline, before starting their work.

Warning and Precautions

Due to chemical material usage that may be hazardous, always make sure to wear suitable lab coat, disposable gloves, and protective eyewear. Material Safety Data Sheet (MSDS) for all products and reagents are provided. They are accessible online at ROJE Technologies

Quality Control

RNall Kit is tested against predetermined experiments on a lot-to-lot basis according to ROJETechnologies ISO-certified quality management system, to ensure consistent product quality. For your information, the results of all experiments are accessible by addressing Cat and Lot number on web at ROJE Technologies

Procedure

The RNall kit is designed for isolating all forms of RNA molecules from 0.5-100 mg tissue samples, up to 1×107 animal or bacteri cells and  1-10 ml of whole blood. Fresh or frozen samples should be lysed and homogenized. For procedure in which  RNsol is used as lysis buffer, Aqueous phase is separated using chloroform. After lysing samples,  the rest of procedure is the same. RNA binding to the silica membrane is achieved selectively, by the addition of Ethanol to the lysate. Contaminants removed by two specific kinds of washing buffers. Pure RNA and miRNA is finally eluted in nuclease-free water. Isolated RNA is ready to use in downstream amplification. It has A 260/ A 280 ratios of 1.9–2.3 by spectrophotometer, confirming high purity.

Equipment & Reagents to be supplied by user

  • Absolute Ethanol
  • 1.5 ml RNase free microcentrifuge tubes
  • RNase free pipets and pipet tips
  • Benchtop refrigerated microcentrifuge (while RNSol is used)
  • Vortex
  • Thermoblock or water bath
  • TissueLyser/ Mortar and pestle (for animal tissue sample)
  • Syringe and needles
  • Homogenizer

Applications

The isolated RNA and miRNA can be used in many downstream applications:

  • RNA Sequencing
  • cDNA synthesis
  • Poly A+ RNA selection
  • Microarrays
  • RT-PCR and real-time RT-PCR
  • Northern, dot blot analysis

Recommended starting material:

To reach optimized results it is better to follow as listed below. The size of the recommended starting material to use with determined RLB, RNSol, Chloroform and Ethanol volumes, are written in Table 2 and 3.

Table 2. Appropriate sample size and amount of RLB

 

Sample Type Sample Size Amount of RLB
Animal Cells ≤5 X106 350 µl
Animal Cells 5 X 106 To 1 X 107 600 µl
Animal Not Fibrous Tissues 0.5-10 Mg 350 µl
Animal Not Fibrous Tissues 10-30 Mg 600 µl
Animal Fibrous Tissues 0.5-30 Mg 300 µl
Bacteria Cells ≤5 X108 500 µl
Bacteria Cells 5 X 108 – 1 X 109 800 µl
Bacteria Cells 1 X 109 – 2 X 109 1000 µl
Whole blood Up to 2 x 106  cells 350 µl
Whole blood 2 x 106 to 1 x 107 cells 600 µl

 

 

Table 3. Appropriate sample size and amount of RNSol Reagent, chloroform and Ethanol amount

 

Sample Size RNSol Reagent

Amount

Chloroform Amount
10 mg animal tissue,

1 × 106 animal cells

100 µl 100 µl
20 mg animal tissue

2 × 106 animal cells

200 µl 100 µl
30 mg animal tissue

3 × 106 animal cells

0.5-1 ml whole blood

300 µl 200 µl
40 mg animal tissue

4 × 106 animal cells

400 µl 200 µl
50 mg animal tissue

5 × 106 animal cells

1-3 ml whole Blood

500 µl 300 µl
60 mg animal tissue

6 × 106 animal cells

600 µl 300 µl
70 mg animal tissue

7 × 106 animal cells

3-5 ml whole Blood

700 µl 400 µl
80 mg animal tissue

8 × 106 animal cells

800 µl 400 µl
90 mg animal tissue

9 × 106 animal cells

900 µl 500 µl
100 mg animal tissue

107 animal cells

5-10 ml whole Blood

1ml 500 µl

Technical Assistance

ROJETechnologies guarantees your complete satisfaction. ROJE technical support team composed of highly trained experienced scientists, who are able to troubleshoot most problems you face. Our technical support team can offer expert advice which may help you select suitable product.

Contact our technical support at any time by selecting one of these ways:

  • Through our telephone and fax number available at ROJETechnologies website.
  • You can submit your question directly to ROJE Technical Support Team from our website (rojetechnologies.com)

Or send your questions to this email address, [email protected]

 

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Additional information

Size

100 prep, 50 prep

Specification

Features Specifications
Elution Volume (µl) 30-100 µl
Technology Silica technology
Main Sample Type Animal cells and tissue /Bacteria cells/PBMC
Processing Manual
Purification of Total RNA, Mirna, Poly A+ Mi-RNA, DNA Or Protein Yes
Sample Amount Varies
Operation Time Per Reaction (Min) Less than 30 min (for RNA Isolation ≥200 nucleotides)

Less than 45 min (for Total RNA Isolation)

Typical Yield (µg) Varies depending on sample type
Average Purity (A260/A280, A260/A230) A260/A280= 1.9-2.3
Size of RNA Purified Total RNA
Enzyme Only for fibrous tissue and bacteria when using RLB (phenol-chlorophorm-free method)

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Protocol

none!

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Resources

MSDS RNall Kit

Hand Book RNall Kit

Quick Protocol EN  FA

Video Protocol Download

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