RNAprep Pure Cell/Bacteria Kit

For purification of high-quality total RNA from cells and bacteria.

The RNAprep Pure Cell/Bacteria Kit provides a fast, simple and cost-effective method for purification of total RNA from cultured cells and bacteria samples by using effective spin column and unique buffer system. The kit includes RNase-Free Spin Column CR3 for purifying high quality RNA by using silica-membrane technology. High-quality total RNA could be obtained in 30-40 minutes with high-purity and is free of protein and genomic DNA contamination.

Cat. No	Packing Size
4992235	50 preps

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Features

■ Optimized buffers and protocols for cultured cells and bacteria samples make the process simple and convenient.
■ Unique DNase I minimizes genomic DNA contamination.
■ Unique RNase-Free Filtration Columns CS eliminates other contaminations.
■ The high-purity and ready-to-use RNA is suitable for sensitive downstream applications.
■ No phenol/chloroform extraction, no LiCl and ethanol precipitation, no CsCl gradient centrifugation are needed, which makes the process safe and reliable.

Applications

■ RT-PCR.
■ Northern Blot, Dot Blot.
■ Real-Time PCR.
■ Chip analysis.
■ PolyA Screening, in vitro translation, RNase protection analysis and molecular cloning.

All the products can be customized for ODM/OEM. For details, please click Customized Service(ODM/OEM)

Experimental Example

	Material: Human Jurkat Cells (1×10⁶ ) Method: The total RNA of Human Jukat Cells was isolated using the RNAprep Pure Cell/Bacteria Kit. Results: Please see the above agarose gel electrophoresis picture. 2-4 μl of 50 μl eluates were loaded per lane. The electrophoresis was conducted at 6 V/cm for 30 min on 1% agarose gel.
	Material: TOP10 E.coli (1×10⁸) Method: The total RNA of TOP10 E.coli was isolated using the RNAprep Pure Cell/Bacteria Kit. Results: Please see the above agarose gel electrophoresis picture. 2-4 μl of 50 μl eluates were loaded per lane. The electrophoresis was conducted at 6 V/cm for 30 min on 1% agarose gel.

FAQ

Q: Column blockage

A-1 Cell lysis or homogenization not sufficient

---- Reduce sample usage, increase the amount of lysis buffer, increase homogenization and lysis time.

A-2 Sample amount is too large

---- Reduce the amount of sample used or increase the amount of lysis buffer.

Q: Low RNA yield

A-1 Insufficient cell lysis or homogenization

---- Reduce sample usage, increase the amount of lysis buffer, increase homogenization and lysis time.

A-2 Sample amount is too large

----Please refer to the maximum processing capacity.

A-3 RNA is not eluted completely from the column

---- After adding RNase-Free water, leave it for a few minutes before centrifuging.

A-4 Ethanol in the eluent

---- After rinsing, centrifuge again and remove the washing buffer as much as possible.

A-5 Cell culture medium is not completely removed

---- When collecting cells, please make sure to remove the culture medium as much as possible.

A-6 The cells stored in RNAstore are not effectively centrifuged

----RNAstore density is greater than the average cell culture medium; so the centrifugal force should be increased. It is suggested to centrifuge at 3000x g.

A-7 Low RNA content and abundance in the sample

---- Use a positive sample to determine if the low-yield is caused by the sample.

Q: RNA degradation

A-1 The material is not fresh

---- Fresh tissues should be stored in liquid nitrogen immediately or immediately put into the RNAstore reagent to ensure the extraction effect.

A-2 Sample amount is too large

---- Reduce sample amount.

A-3 RNase contamination

----Although the buffer provided in the kit does not contain RNase, it is easy to contaminate RNase during extraction process and should be handled with care.

A-4 Electrophoresis pollution

---- Replace the electrophoresis buffer and make sure the consumables and Loading Buffer are free of RNase contamination.

A-5 Too much loading for electrophoresis

---- Reduce the amount of sample loading, the loading of each well should not exceed 2 μg.

Q: DNA contamination

A-1 Sample amount is too large

---- Reduce sample amount.

A-2 Some samples have high DNA content and can be treated with DNase.

----Perform RNase-Free DNase treatment to the obtained RNA solution, and the RNA can be directly used for subsequent experiments after treatment, or can be further purified by RNA purification kits.

Q: How to remove RNase from experimental consumables and glasswares?

For glasswares, baked at 150°C for 4 h. For plastic containers, immersed in 0.5 M NaOH for 10 min, then thoroughly rinsed with RNase-free water and then sterilize to completely remove RNase. The reagents or solutions used in the experiment, especially water, must be free of RNase. Use RNase-free water for all reagent preparations (add water to a clean glass bottle, add DEPC to a final concentration of 0.1% (V/V), shake overnight and autoclave).

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