What is the best way to remove/disolve spawn jelly in order to extract DNA/RNA/proteins from snail eggs with TRIZOL?

I'm interested in the shape and size of the radula of Radix species. Especially the dimensions of the single teeth. Also traces of radula scratches would be helpful.

I'm looking to try and extract nucleic acid from FTA cards, has anyone had trizol extraction to work well?

Thanks

Hi, I am preparing a new project on lncRNAs in human cancer cell lines. I scanned through many high-impacted journals to find the best RNA isolation kit that would also isolate small RNAs and that would be compatible with RNA-Seq. I noticed that pretty much everyone is using Trizol reagent for RNA extraction, followed by column-based purification, with three kits being the most popular: RNAEasy kit, miRVana kit or Trizol Plus kit (Trizol together with purification columns). Which column-based purification system is the best one, in terms of RNA purity and stability and good quality for subsequent lncRNA RNA-Seq, with possibility to run also small RNA-Seq for miRNAs in the future?

I need to dissect the the reproductive system from the snails but can't find any information/ protocol on how to dissect this species. Can anyone point me in the direction of somewhere that may have this information?

I have high peaks in the fast region of my total RNA as seen in the attached pictures. The samples 9 and 12 have different problems, with 12 probably being somehow contaminated and 9 being degraded. But the remaining samples show the very high peaks around 25- 30s which correspond to where I would expect small RNAs and 5s RNA. But they are much bigger than my rRNA peaks. What could be the cause of these and will they pose a problem in downstream applications?

Extraction was done using trizol from gram negative bacteria.

I know that trizol yields a higher proportion of small RNAs then other extraction methods, but this seems like a lot to me.

Thank you for your answers!

Biology and ecology that means the growth, reproduction, resistance to the negative environmental factors (elevated temperature, anthropogenic contamination, etc., and so on.

I would be very appreciated for it.

Sincerely yours,   

prof. A.Golubev

Hi there,

I know that there are very good kits for miRNA isolation, but I want to use Trizol for this purpose. It is known that Trizol selectively loses GC-rich small RNAs when extracting RNA from small starting material, and I guess it may still have this same problem when using larger starting material for RNA isolation (i.e.losing a subset of miRNAs).

Does anyone of you have specific comments on how to successfully and efficiently use Trizol for total RNA isolation which contains small RNAs as well? What modifications should be applied to Trizol isolation so that small RNA could be preserved?

Thank you in advance,

Sharif

Hello, 

I am about to IP my protein of interest and maybe (I hope so) some small RNA would co-IP with it. Since this is my first time in this area, would anyone be so kind to suggest the best method for separating these RNAs for the further analysis (based on your own experience [good & bad])? I am thinking of trizol but not really sure.

Thank you in advance!

Dear Folks,

Greetings. Recently I am isolating total RNA as well as small RNA for small RNA-seq. I am using trizol based kit "RNAzol-RT (Cat-RN190). I am facing big problem regarding with quality as well as quantity. Could anyone plzz help me from their experience with tomato seeds?

Here is the brief protocol I used for RNA isolation-

1. 1ml RNAzol+100mg seed powder

2. Add 400ul water and incubate 15 mins followed by 12000gx15 min centrifuse

3. 1ml suppernatent + 400ul of 75% ethanol>10 min incubation>12000gX8 min centrifuse

small RNA precipitation:

4. 1 ml suppernatent + 0.8volume of isopropanol > incubate 30 min> 12000gx 15 mins

5. wash the pellete two times with 70% isopropanol 8000gx3 min

6. disolve the pellete with RNAse free water.

With this protocol I really got both poor RNA quality and concentration.

I would again request if anyone can share their practicle experience that will be great help or me..

Best

Torikul Islam PhD