The study of the molecular processes of the nucleus and cell function.
3
votes
1answer
107 views
Why re-label the strand ends in 3' DNA labelling?
I have a problem with a molecular biology question; I don’t understand how DNA 3’ labelling works.
I took a diagram from my lesson and tried to understand with it; this is what I understood.
If I’m ...
2
votes
1answer
51 views
Histone marks mechanism
I am slightly confused about the mechanisms that makes histone modifications associate with gene expression.
That is, H3K36me3 is believed to be present in actively transcribed genes, H3K27me3 in ...
6
votes
0answers
47 views
Does the MS2 RNA binding protein have any translational repression effects?
Reposted from Quora: http://www.quora.com/Does-the-MS2-RNA-binding-protein-have-any-translational-repression-effects
I'm thinking of the MS2 protein binding to its RNA hairpin target. Would the ...
5
votes
0answers
61 views
What is the fastest way to build an alanine scanning library?
For interfacial studies, I would like to build an alanine scanning library for one of my proteins examining 20 sites. I will ultimately express the gene using E.coli cell-free protein synthesis. I ...
3
votes
0answers
25 views
How does LCR compare to Assembly PCR
The question pretty much explains itself. How do the two methods compare? I've always used Assembly PCR but the method is prone to mistakes and I'm curious how it compares to Ligase Chain Reaction ...
2
votes
0answers
37 views
What is the purpose of Y-shaped adapters in Illumina sequencing?
Y adapters different sequences to be annealed to the 5' and 3' ends of each molecule in a library.
The arms of the Y are unique, and the middle part, connected to the DNA fragment, is complementary.
...
2
votes
0answers
58 views
Optimization of annealing temperature
I am trying to optimize annealing temperature for some primer pairs. I have tried optimization using cDNA, genomic DNA, Taq polymerase, phusion polymerase etc., but every time I am either getting ...
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?
...
1
vote
0answers
34 views
Low RNA yield, low 260/230 ratio
I extracted total RNA from animal tissue using the Qiagen RNeasy kit, however my RNA yield was extremely low and the 260/230 ratio was around 0.3. This is the protocol I followed:
The animal ...
1
vote
0answers
26 views
What mammalian cell lines are easiest to grow and transfect?
I'm working with HEK293 cells, which are very easy to transfect. Does anyone have other suggestions for cell lines with very high transfection efficiency? Ideally the chromosomes would also be diploid ...
1
vote
0answers
35 views
Can I heat Trizol?
I wonder if I can heat Trizol reagent for 30 min 65C. The goal is to disrupt protein-RNA complex while inhibiting nucleases. (I can't use RNasin cause it's inactivated in 65C, and can't use RVC cause ...
1
vote
0answers
28 views
Experience with Detox Cloning
I just saw this paper.
A Highly Efficient Molecular Cloning Platform that Utilises a Small Bacterial Toxin Gene
I was curious if anyone had experience with this cloning strategy using the IbsC ...
1
vote
0answers
53 views
Molecular cause of cramps, spasms and strengthening in muscles? (incl. intro to muscle contraction)
When motor neurons are stimulated to trigger an action potential, this potential propagates down the spine, eventually reaching a neuromuscular junction, causing the release of acetylcholine (ACh).
...
1
vote
0answers
44 views
Solid phase use in HIV rapid tests
I have another question in regards to my HIV test research. The rapid tests like Orasures Oraquick contains a strip of synthetic peptides that are used to represent proteins found in the envelope ...
0
votes
0answers
35 views
Hardy-Weinberg applied to three alleles and stimation of allele frequencies
I have this equation:
Corresponds to HW in equilibria with three alleles:
$(p+q+r)^2=1$
Expanding the square results:
$p^2+2pq+r^2+2pr+q^2+2qr = 1$
I need to separate homozygous and ...
