Dear Bhaumik Patel

You can use TRIzol® Reagent

Is a complete, ready-to-use reagent for the isolation of high-quality total RNA or the simultaneous isolation of RNA, DNA, and protein from a variety of biological samples. This monophasic solution of phenol and guanidine isothiocyanate is designed to isolate separate fractions of RNA, DNA, and proteins from cell and tissue samples of human, animal, plant, yeast, or bacterial origin, within one hour.

Key features of TRIzol® Reagent include:

• Permits the isolation of RNA, DNA, and protein from the same sample

• Offers superior lysis capability, even with difficult sample types

• Optimized formulations and protocols for tissues, cells, serum, virus, and bacteria

Reliably purify RNA from multiple sample volumes and sources

TRIzol® Reagent performs well with small quantities of tissue (50–100 mg) and cells (5 × 106), as well as with large quantities of tissue (≥1 g) and cells (>107), and comes with prototcols for purification from samples of human, animal, plant, or bacterial origin. TRIzol® Reagent maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization. The simplicity of the TRIzol® Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in 1 hour. Total RNA isolated by TRIzol® Reagent is free of protein and DNA contamination.

Formulated for isolation of multiple molecular targets

TRIzol® Reagent allows you to perform sequential precipitation of RNA, DNA, and proteins from a single sample. After homogenizing the sample with TRIzol® Reagent, chloroform is added, and the homogenate is allowed to separate into a clear upper aqueous layer (containing RNA), and interphase and red lower organic layers (containing the DNA and proteins). RNA is precipitated from the aqueous layer with isopropanol. DNA is precipitated from the interphase/organic layer with ethanol. Protein is precipitated from the phenol-ethanol supernatant by isopropanol precipitation. The precipitated RNA, DNA, or protein is washed to remove impurities, and then resuspended for use in downstream applications.

It is not a pleasant answer but in your case you should not expect to get a good quality RNA or having right expression profiles from the samples. To get stable RNA the cells must be broken or the RNA must be present in the buffer where the enzymes could be suppressed which could lyse RNA. Nevertheless, you can try products like RNAstable, but it also cant ensure the quality. For DNA storage same principle applies, but as DNA is very stable molecule and less prone to degradation a short term storage in buffer is okey. You can find more answers and suggestions here. https://www.researchgate.net/post/Best_long_term_storage_method_for_DNA

Hello,

I agree with Abhijeet, for RNA isolation fresh samples should be immediatelly inserted into special solution (RNAlater, RNA stable,...) and after app. 1 hour (solution must penetrate into sample) frozen at -20°C.

https://www.thermofisher.com/cz/en/home/brands/product-brand/rnalater.html

Dear patel:- I could say NAsafe solution is the best as follow:-

NAsafe is salting-out all proteins media of ammonium sulfate (the saturating concentration) with pH < 5 (see the lyotropic series).

The description of NAsafe (4 M (NH4)2SO4, 10 mM EDTA, 0.1 M Mes, pH 4.6):

combine 40 ml 0.5 M EDTA, 19.5 g Mes free acid, 528 g ammonium sulfate, dissolve in Milli-Q water, bring final volume 1L; stir on a hot plate stirrer on low heat until the ammonium sulfate is completely dissolved. Transfer to a screw top bottle and store either at room temperature or refrigerated.

Store NAsafe solution at room temperature. It is guaranteed for 6 months from the date of receipt, if properly stored. If any precipitation of NAsafe solution is seen, heat it to 37°C and agitate to redissolve it.

RNase inactivation is reversible; do not rinse NAsafe solution from samples before using. Blot tissues with a wipe, or pellet cells to remove excess NAsafe solution..

The guideline for use NAsafe:-

• Use NAsafe solution with fresh tissue only; do not freeze tissues before immersion in NAsafe solution.

• Before immersion in NAsafe solution, cut large tissue samples to ≤ 0.5 cm in any single dimension.

• Place the fresh tissue in 5–10 volumes of NAsafe solution.

• Most samples in NAsafe solution can be stored at room temp for 1 week without compromising RNA quality, or at –20°C or –80°C indefinitely.

• Do not freeze samples in NAsafe solution immediately; store at 4°C overnight (to allow the solution to thoroughly penetrate the tissue), remove supernatant, then move to –20°C or –80°C for long-term storage.

Most samples can be stored at 25°C in NAsafe solution for up to 1 week without significant loss of RNA quality. After 2 weeks at 25°C, RNA generally appears slightly degraded (marginally acceptable for Northern analysis, but still of sufficient quality for nuclease protection assays or RT-PCR analysis).

Hope I can answer your question accurately...Regard

Thanks Abhijeet and Martin for the valuable information but I am only going to get the sample after 3 days. maximum they can send me the sample on the ice (& may be not always possible) so I need the preservation buffer to get the good quality of the RNA for the consistency of the sample.

Thanks Dr. Mushtak T. S. Al-Ouqaili for the giving the recipe of the NAsafe solution with the guidelines. I will use it for my tissue in coming day. You already mentioned that it is for the tissue but I am curious that does it also work with the whole blood sample collected in the EDTA tube ? I can transfer small amount of the blood (EDTA) to NAsafe solution like the RNAlater. If it can, then I think i can use it universally(tissue, hair and blood).

Please give me your suggestions.

Thanks -Bhaumik

To my view no single liquid method is suitable for preservation.The choice to be based on experimental design, field facilities, shipping constraints etc.

Dear Bhaumik Patel

You can use TRIzol® Reagent

Is a complete, ready-to-use reagent for the isolation of high-quality total RNA or the simultaneous isolation of RNA, DNA, and protein from a variety of biological samples. This monophasic solution of phenol and guanidine isothiocyanate is designed to isolate separate fractions of RNA, DNA, and proteins from cell and tissue samples of human, animal, plant, yeast, or bacterial origin, within one hour.

Key features of TRIzol® Reagent include:

• Permits the isolation of RNA, DNA, and protein from the same sample

• Offers superior lysis capability, even with difficult sample types

• Optimized formulations and protocols for tissues, cells, serum, virus, and bacteria

Reliably purify RNA from multiple sample volumes and sources

TRIzol® Reagent performs well with small quantities of tissue (50–100 mg) and cells (5 × 106), as well as with large quantities of tissue (≥1 g) and cells (>107), and comes with prototcols for purification from samples of human, animal, plant, or bacterial origin. TRIzol® Reagent maintains the integrity of the RNA due to highly effective inhibition of RNase activity while disrupting cells and dissolving cell components during sample homogenization. The simplicity of the TRIzol® Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in 1 hour. Total RNA isolated by TRIzol® Reagent is free of protein and DNA contamination.

Formulated for isolation of multiple molecular targets

TRIzol® Reagent allows you to perform sequential precipitation of RNA, DNA, and proteins from a single sample. After homogenizing the sample with TRIzol® Reagent, chloroform is added, and the homogenate is allowed to separate into a clear upper aqueous layer (containing RNA), and interphase and red lower organic layers (containing the DNA and proteins). RNA is precipitated from the aqueous layer with isopropanol. DNA is precipitated from the interphase/organic layer with ethanol. Protein is precipitated from the phenol-ethanol supernatant by isopropanol precipitation. The precipitated RNA, DNA, or protein is washed to remove impurities, and then resuspended for use in downstream applications.

agree with @ Mushtak T. S. Al-Ouqaili

regards

Also,,

Kindly follow link below for some suggestions

https://www.researchgate.net/post/Isolation_of_RNA_and_DNA_from_blood_stored_in_EDTA_vacutainer

I wish you good luck.

Thanks for the suggestions. Mohammed H. Musleh.