hello,
I am extracting RNA from Blood and saliva using Promega Reliaprep RNA cell miniprep system
in the lysis step it says that this step is to lyse cultured cells from suspension or adherent cultures however my samples are stains of blood and saliva sputum, many researchers have used this kit with the same form of samples as mine and got RNA, in my case i can't get RNA, no bands on the gel!!! any idea?
Yes, I don't think it matters, the kit should work well. Most important thing with RNA is to get a fresh sample and process it quickly. Storing them in RNA later or sinking them in the trizol, trizol-like reagent.
for good result must be used fresh sample and if concentration low transfer RNA TO
cDNA
Hi Dr.RNA extraction is the purification of RNA from biological samples. This procedure is complicated by the ubiquitous presence of ribonuclease enzymes in cells and tissues, which can rapidly degrade RNA. Several methods are used in molecular biology to isolate RNA from samples, the most common of these is Guanidinium thiocyanate-phenol-chloroform extraction. The filter paper based lysis and elution method features high throughput capacity. RNA extraction in liquid nitrogen, commonly using a mortar and pestle (or specialized steel devices known as tissue pulverizers) is also useful in preventing ribonuclease
Dear sir
You can RNA extraction in liquid nitrogen, commonly using a mortar and pestle (or specialized steel devices known as tissue pulverizers) is also useful in preventing ribonuclease
Hello,
you can use miRNA easy kit is very good for the PBMC (pellet of cells of mononuclar cells)
https://www.qiagen.com/de/shop/sample-technologies/rna/mirna/mirneasy-mini-kit/#orderinginformation
I had a very good Quality of rna in bioanayser and I had around 2ug to 15 ug of RNA in total depending to the Initial amount of material.
otherwise
you can use the Lysis buffer this kit Qiam to remove the red cells and then use the miRNA easy kit for the next steps
https://www.qiagen.com/de/shop/sample-technologies/rna/total-rna/qiaamp-rna-blood-mini-kit/#orderinginformation
hi dr. nono pleas read this artical
Article Evaluation of Existing Methods for Human Blood mRNA Isolatio...
Hello,
I used this kit for total intracellular RNA extraction from fresh blood treated with UFH:
E.Z.N.A Blood RNA Kit (VWR, Omege bio - tek).
It worked perfectly with freshly collected blood, and from blood stored at 4 °C for 2 days, as well.
Also, check if the problem is not in the reverse transcription. I used Super script III Reverse Transcriptase from Invitrogen.
One of my friends faced the same issue too. However, he continued with cDNA conversion and qPCR too and finally he got it.
The issue here was too small amt of RNA. You may try to continue with what you supposed to do. See how first. At the same time, try to modify the way you extracted RNA or use different kit and fresh samples as suggested.
Hello,
For the blood RNA, I recommend you use ReliaPrep RNA Tissue Miniprep system (Z6112).
This chemistry would be more effective than RP RNA cell kit if you apply the buffy coat into the kit to extract RNA from whole blood.
Regarding the saliva (Oragen?), did you try to incubate at 50ºC for hours?
If you did, I'd like to recommend you remove the sputum as possible as you can after incubation.
Please let me know if you need the detail instruction.
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