RNall Kit

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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