Tagged Questions
2
votes
3answers
63 views
Knockdown of long noncoding RNA (lncRNA) - how is it done?
I don't work at the wet lab and don't know all the details about the knockdown techniques.
My question is:
How lncRNA knockdown is done?
For example - you have lncRNA that is functional in the ...
2
votes
1answer
39 views
Does bleach destroy RNAse activity, and if so, how does it do it?
I am working with RNA samples, and I'm trying to be very careful about RNAse contamination.
I have some questions about bleach, though. Some people say that a solution of bleach is enough to destroy ...
5
votes
2answers
59 views
Is wiping with RNAse Zap enough to destroy RNAse activity?
From the RNAseZap MSDS, it is an SDS at some unknown concentration, maybe with some NaOH?
Some other links suggest there is some NaOH as well.
The Ambion site states that RNAseZap destroys RNAse ...
5
votes
1answer
199 views
Very high 260/230 absorbance ratio of an RNA sample
After my most recent RNA extraction, the RNA samples had very high 260/230 absorbance ratios, (ranging from 5 to 25).
I've never gotten numbers like this and I know the ratio is supposed to be ~2 in ...
5
votes
1answer
696 views
What is the function of the RNA primer in DNA replication?
During DNA replication, RNA primase puts an RNA primer in the lagging strand. What is the function of this RNA primer? Why can't the enzymes put DNA fragments directly?
6
votes
1answer
37 views
circulating microRNAs are functional?
In plasma and other body fluids, miRNAs can be found. They not only originate from dying cells but also from active secretion and are usually 'packed' into vesicles/lipo-proteic structures (i.e. ...
6
votes
1answer
551 views
Purpose of poly(A)+ RNA?
I am learning RNA-seq analysis. I always encounter this phase "poly(A)+ RNA". After searching, I got this: "Most messenger RNAs contain a poly(A) tail, while structural RNAs do not. Poly(A) selection ...
2
votes
0answers
73 views
Mouse meta-globin mRNA problem
This is an mRNA strand of mouse meta-globin:
5'-ccccagauacggaauucgaau-3'
A) Which small RNA (below) is most likely to regulate expression of
meta-globin?
...
15
votes
2answers
392 views
How long can I store extracted RNA?
If I extract RNA from a (leaf tissue) sample using a one-step phenol:chloroform extraction, how long can those samples be stored at -80°C? And how many times can I defrost and refreeze them before ...
10
votes
2answers
2k views
How do I clean phenol contaminated RNA without losing any of the sample?
I recently extracted RNA from developing plant leaves for the first time, as part of a very long and intensive experiment. The samples were extremely precious because of the amount of effort that went ...
6
votes
2answers
443 views
Reverse transcription PCR optimization
What is the ideal amount of RNA to use for the RT? and how much cDNA to use then for the PCR?
I did RT with a solution of RNA of 0.36 ug/ul. Then for my PCR I used 1 ul of the cDNA obtained and used ...
8
votes
1answer
10k views
Absorption ratios 260/280 and 260/230 for RNA
I extracted RNA from different cell lines, an I want to perform reverse transcription and then PCR. To get good results, in which range should the absorption ratios 260/280 nm and 260/230 nm be? And ...
