Isolation RNA from whole blood by using ZR Whole-Blood RNA MiniPrep™

The ZR Whole-Blood RNA MiniPrep™ provides a streamlined method for the rapid isolation of total RNA from whole or partitioned blood in as little as 10 minutes.
RNA can be isolated immediately from fresh samples or at a later time from blood stored in Blood RNA Buffer.  This product is designed to isolate RNA from blood for subsequent RNA-based methods including RT-PCR, hybridization, etc.

The Flowchart

source: zymo

Epigenetic: RNA methylation by using EZ RNA Methylation Kit

What is Epigenetic?

Biological mechanisms in which DNA, RNA, and proteins are chemically or structurally modified, without changing their primary sequence (a.k.a mutation). 

Epigenetic regulatory mechanisms include DNA methylation and hydroxymethylation, histone modification, chromatin remodeling, RNA methylation, and regulation by small and long non-coding RNAs.

What is RNA methylation?

 Although the majority of nucleic acid modification research involves 5-methylcytosine (5-mC) in DNA, RNA is also extensively modified. In fact, there exist over a hundred modifications to RNA, including 5-mC, and RNA methylation is a common and naturally-occurring event in both prokaryotic and eukaryotic organisms. RNA methylation has been observed in numerous types of RNA molecules, including mRNAs, tRNAs, and non-coding RNAs, but the function of RNA methylation remains unclear.

Source: Zymo

How to use EZ RNA Methylation Kit ?

Follow the following flowchart and the complete protocol here

Source: Zymo

What happened to RNA methylation?

Sequencing results following bisulfite treatment. RNA with methylated C (5-mC) at nucleotide position #7 was processed using the EZ RNA Methylation™ Kit. The recovered RNA was amplified by RT-PCR and then cloned and sequenced. The methylated cytosine at position #7 remained intact while the non-methylated cytosines at positions #3, 4, and 13 were completely converted into uracil (post-bisulfite treatment) and detected as thymine following RT-PCR and sequencing

 

 Source: Zymo

Epigenetic: bisulfite treatment using Zymo kit (DNA methylation)

What is Epigenetic?

Biological mechanisms in which DNA, RNA, and proteins are chemically or structurally modified, without changing their primary sequence (a.k.a mutation). 

Epigenetic regulatory mechanisms include DNA methylation and hydroxymethylation, histone modification, chromatin remodeling, RNA methylation, and regulation by small and long non-coding RNAs.

Is it normal for our body to have DNA methylation?

 Yes, it is normal for our body, in fact recently many diseases due to aberration in epigenetic function. 

In complicated terms these epigenetic modifications normally play critical roles in the regulation of numerous cellular processes, including gene expression, DNA replication, and recombination

Source: Zymo

What kind of disease due to epigenetic aberration?  

 Well the most obvious is cancer.

Source: uiuc.edu

 

What is DNA methylation?

DNA which has methyl group (-CH3) on its nucleotide (especially Cytosine or C in mammals)

Source: uiuc.edu

Or in complicated term DNA methylation: Methylation of cytosine is a covalent modification of DNA, in which hydrogen H5 of cytosine is replaced by a methyl group 

Source: Sigma Aldrich

 

What is bisulfite treatment?

Converting C to be U by using sodium metabisulfite

What is bisulfite protocol?

What kind of kit to choose?

Zymo Research has several kit depend on your need, the following is recommended to see.

Source:
Zymo Research