MiRJia PB Kit,(with RBC Lysis Buffer,1X)

Description

Kit content

Component	50 preps	100 preps
RNSol Reagent	50 ml	100 ml
TWB1 (concentrate)	17 ml	35 ml
TWB2 (concentrate)	15 ml	27.5 ml
Nuclease free RBC Lysis Buffer Just supplied in MiRJia PB Kit with RBC Lysis Buffer	2.5 L	5 L
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 and nuclease free RBC Lysis buffer 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 MiRJia PB Kit provides the procedures necessary for purifying total RNA from peripheral blood mononuclear cell. Notice that, MiRJia PB Kit 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 rojetechnologies.com

Quality Control

MiRJia PB 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 rojetechnoloes.com

Procedure

The MiRJia PB Kit is designed for isolating all forms of RNA molecules from up to 1×10⁷ PBMCs. Fresh or frozen samples should be lysed and homogenized. Aqueous phase is separated using chloroform. Then, RNA binding to the silica membrane is achieved selectively, by the addition of Ethanol to the lysate. Contaminants are removed by two specific kind of washing buffers. Pure miRNA and total RNA is finally eluted in nuclease free water. Isolated RNA (miRNA or total RNA) is ready to use in downstream applications. 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
• Pipets and RNase free pipet tips
• Benchtop refrigerated microcentrifuge
• Vortex
• TissueLyser/ Mortar and pestle/ Homogenizer
• Syringe and needles

Applications

The isolated RNA can be used in many downstream applications:

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

Recommended starting material:

To reach optimized results it is better to follow as listed below. The size of the recommended material to use with determined RNSol and chloroform amount are written in Table 2.

Table 2. Appropriate sample size and amount of RNSol, Chloroform and Ethanol amount

Whole blood	RNSol Amount	Chloroform Amount	Ethanol Amount
1 ml to 3 ml	500 µl	300 µl	400 µl
3 ml to 5 ml	700 µl	400 µl	400 µl
5 ml to 10 ml	1 ml	500 µl	400 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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