TRzol (Total RNA Extraction Reagent)

TRzol (Total RNA Extraction Reagent)

Product Introduction

TRzol reagent is a reagent that directly extracts total RNA from cells or tissues. It can maintain the integrity of RNA when breaking and dissolving cells. After adding chloroform and centrifuging, the sample was divided into a water sample layer and an organic layer. RNA exists in the aqueous layer. After collecting the water sample layer above, RNA can be recovered by isopropanol precipitation.

This method has good separation effects on small and large amounts of tissues and cells, whether they are human, animal, plant, or bacterial tissues. The simplicity of TRzol reagent operation allows for the simultaneous processing of multiple samples. All operations can be completed within one hour. The total RNA extracted by TRzol can avoid contamination of DNA and proteins. Therefore, it can perform RNA blot analysis, spot hybridization, poly (A)+selection, in vitro transcription, RNAse protection analysis, and molecular cloning.

TRzol reagent can promote the precipitation of various RNAs of different species and molecular weights. For example, RNA extracted from rat liver was electrophoretic on agarose gel and stained with ethidium bromide, and many discontinuous high molecular weight bands (mRNA and hnRNA components) between 7 kb and 15 kb, two dominant ribosomes~5kb (28S) and~2kb (18S), and low molecular weight RNA between 0.1 and 0.3 kb (tRNA, 5S) were seen. When the extracted RNA is diluted with TE, its A 260/A 280 ratio is ≥ 1.8.

Product storage: TRzol can be stably stored for 12 months at room temperature. However, for optimal results, we recommend storing in an environment of 2-8 ℃. (For specific instructions, please refer to the manual)

1、 Template DNA: PCR reaction requires template DNA, such as extracting DNA from cells, tissues and blood;

2、 Primers: Two primers used in PCR reactions that pair with specific regions required for amplification at both ends of the template DNA. Primers can also be synthesized into primers used in processes such as TA cloning;

3、 DNTPs: Four types of dNTPs are required in PCR reactions, including deoxyadenosine monophosphate (dATP), deoxythymidine monophosphate (dCTP), deoxyguanosine monophosphate (dGTP), and deoxycytosine monophosphate (dTTP), which are used in biological processes such as cell division and DNA synthesis, as well as for the extension and amplification of template DNA chains in PCR amplification;

4、 Taq DNA polymerase: a polymerase required for PCR reactions, generally using Taq polymerase, as well as other types of polymerases such as PFU, Phusion, etc., for amplifying DNA strands;

5、 PCR buffer solution: It has a buffering effect on the PCR reaction, adjusts the reaction pH, and can enhance the specificity of the PCR reaction by using hydrogen thiosulfate SDS and small molecule organic compounds such as formaldehyde, thereby improving reaction efficiency;

6、 Enzyme digestion system: PCR amplification often involves experimental steps such as recombination, construction, and sequencing, and often requires enzyme digestion operations.

7、 MARK: A molecular weight standard used to determine the size of DNA fragments.

8、 DNA Purification Kit: Used for purifying PCR reaction products;

Nak porridge; oligo DT
When choosing Oligo dT, it is required that RNA must have Poly A, so mRNA from eukaryotes is applicable. RT suitable for long or even full-length mRNA requires high quality RNA samples, without obvious DNA contamination, RNA degradation, and RNA breakage. If you want to explore new mRNA for RT reaction, it is recommended to use Oligo dT primers. The stability of using Oligo dT primers is better than that of random primers and specific primers.
② Random Primers
Suitable for RT of various RNAs, especially for situations with low template abundance (such as low expression of a certain gene). When selecting random primers, all RNA is used as templates in the chain cDNA synthesis reaction, and primers are designed for specific amplification during PCR reaction. At the same time, attention should be paid to the relationship between the amount of random primers and the total RNA amount. It is generally recommended to use 50ng of random primers per 5 µ g of total RNA. If the amount of random primers per 5 µ g of total RNA exceeds 250ng, it may lead to an increase in small fragment products (<500bp) and a decrease in long fragment and full-length products.
③ Specific primers
Gene specific primers (GSP) are complementary primers to a template that are a part of a gene sequence. This primer needs to be designed in combination with the known sequence of the target RNA. Due to the specific binding between primers and RNA sequences, each target RNA requires a new set of gene specific primers. Therefore, when analyzing multiple targets, it is necessary to apply more RNA samples. Moreover, the annealing temperature of different primers is inherently different, so it is generally not recommended to use them. If specific primers are needed, it is recommended to optimize the annealing temperature.