Search found 81 matches

by roniadam
Sun Aug 11, 2013 2:43 am
Forum: Molecular Biology
Topic: Transcrption factor-promoter (cis-element) interaction
Replies: 3
Views: 2944

Transcrption factor-promoter (cis-element) interaction

I want to transfect my cells with a vector for promoter activity analaysis, I need to detect the binding effeciency of some transcription factors to the promoter, what technique should I use? I have read about ChIP but don't know how to design and perform this experiment practically , your help is h...
by roniadam
Fri Jun 01, 2012 6:20 am
Forum: Molecular Biology
Topic: Relative quantitation qRT-PCR
Replies: 0
Views: 1066

Relative quantitation qRT-PCR

Hi

I am using Stepone plus (applied biosystem) to perform a gene expression assay using relative quantitation. My query is regarding the output result. There are 4 types of graph output: linear, Log10, Ln, Log2. What is the best graph type to show in my report??
by roniadam
Sun Apr 29, 2012 9:19 am
Forum: Molecular Biology
Topic: Regulatory elements inside introns
Replies: 3
Views: 3674

Regulatory elements inside introns

Hi I am wondering if intron have regulatory elements other than splicing recognition signals (Donor, Acceptor and branch point sites). because I have found a mutation in the middle of the intron of my gene and noticed that the patient has some phenotypic resulted from this mutation. So is there any ...
by roniadam
Fri Dec 02, 2011 8:30 am
Forum: Bioinformatics
Topic: Transcription factor signals software
Replies: 0
Views: 1762

Transcription factor signals software

Hi

I have a DNA sequence and I need to analyze this sequence by identifying the regulatory elements found in introns. Is there any software that can predict important DNA signals founf in the internal introns?
by roniadam
Wed Apr 27, 2011 4:01 am
Forum: Molecular Biology
Topic: Separation of double stranded DNA
Replies: 7
Views: 2509

By mutagenesis kit (Quikchange II kit). It has been done before with another group and they succeeded in insertion but I was not. I emailed them but there was no reply that is why I posted this problem in this forum.... :(
by roniadam
Tue Apr 26, 2011 4:37 am
Forum: Molecular Biology
Topic: Separation of double stranded DNA
Replies: 7
Views: 2509

Hi Jackbean

This is a piece of PCR product which is called megaprimer because I want to insert this piece into my vector.
by roniadam
Mon Apr 25, 2011 1:53 pm
Forum: Molecular Biology
Topic: Separation of double stranded DNA
Replies: 7
Views: 2509

Separation of double stranded DNA

Hi everyone: I am interested to make a large insertion (780bp) in my vector. This 780 bp is a PCR product (megaprimer), I have used quikchange II mutagenesis kit but couldn't success?? any idea about how to success??? My thinking about the failing in this insertion is in the megaprimer ??? Any sugge...
by roniadam
Fri Oct 15, 2010 6:47 am
Forum: Molecular Biology
Topic: Real time PCR for transcripts analysis
Replies: 2
Views: 1323

Real time PCR for transcripts analysis

Hello

I am using Real time PCR for gene expression analysis (cDNA-PCR), can anyone refer me to a reference article with similar work.
by roniadam
Fri Oct 15, 2010 6:45 am
Forum: Molecular Biology
Topic: DNA- CAP site mutation
Replies: 2
Views: 1833

Re: DNA- CAP site mutation

No .. I mean CAP site at the RNA LEVEL it is either transcription or translation initiation site. So any one can help???
by roniadam
Thu Oct 14, 2010 3:55 am
Forum: Molecular Biology
Topic: DNA- CAP site mutation
Replies: 2
Views: 1833

DNA- CAP site mutation

Hi Everyone

I got confused with the meaning of CAP site.... is it transcription initiation site OR translation initiation site???
by roniadam
Tue Jun 22, 2010 3:48 am
Forum: Molecular Biology
Topic: Transcripts analysis
Replies: 4
Views: 1991

Re: Transcripts analysis

I used T-A cloning vector and for the promotor, I used the gene's itself promotor and it showed a nice expression with the mutant but less expression with the wild type. the mutation was a point mutation (single nucleotide substitusion). Any idea for how to make the wild type to be more expressed th...
by roniadam
Fri Jun 18, 2010 2:39 pm
Forum: Molecular Biology
Topic: Transcripts analysis
Replies: 4
Views: 1991

Transcripts analysis

Hi I am working on in vitro expression analysis. I have transfected a mammalian cell line with a gene of wild type and mutant one. surprisingly, I found that the transcript level of the wild type gene was less than the mutant (which is against the logic theory where the wild type should be expressed...