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micro RNA

29 Apr 2013

Posted by smktmaran in Molecular Biology
while submitting tissue samples for miRNA profilling by next-gen deep sequencing, to have a check or control on the reliability of results is it advisable to add oligos (reverse complimentary sequence of any known distantly/unrelated miRNA sequences)? kindly help me out. Thanks in advance.

110 Views · 1 Replies ( Last reply by pcrman )

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Bioinformatic tools for novel miRNA target prediction

10 Sep 2011

Posted by hamid in Bioinformatics and Biostatistics
Hi
   I am looking for some means to have  a hint about the targets of some unknown micro RNAs. Are there any bioinformatic tools that can predict the target of an unknown micro RNA? The micro RNA sequences in my hand are not related to known classes of miRNAs. My experimental organism is Drosophila. Any suggestions will be greatly appreciated

1,008 Views · 2 Replies ( Last reply by pcrman )

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GHR gene location of water buffalo?

19 May 2013

Posted by iamleos in Genetics and Genomics
i need to know that in bovine and water buffalo the disorders and diseases related to growth hormone receptor (GHR) or growth hormone binding protein (GHBP)?
also the gene location of GHR of water buffalo with refrences.
pleases can anybody help me out?​

53 Views · 1 Replies ( Last reply by pcrman )

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"Dear" GPCR

20 May 2013

Posted by Tom M in Cell Biology
A colleague of mine determined through a genomic array that diseased renal tissue expressed 100x more DNA than non-diseased renal tissue for a G protein coupled receptor that was identified as the dual endothelin 1/angiotensin II receptor (DEAR).  Does anyone know anything about this receptor or can suggest a publication or two that discuss it?  Thanks.

Tom

50 Views · 1 Replies ( Last reply by pcrman )

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Reproducible Non-Specific PCR Product

20 May 2013

Posted by Epigeneticist in PCR, RT-PCR and Real-Time PCR
I am genotyping mouse cells after cre-mediated knockout. The gene has LoxP sites with intron 1 and intron 10, so a large portion of the gene is deleted. To detect deletion by PCR, forward primer anneals within intron 1 and reverse intron 10, which gives a 600bp product. However, I also get a non-specific product. Genomic DNA from control  (floxed sample, no knockout) and gene knockout samples both produce a product that is around 660bp. I sequenced the PCR product and the results are confusing (gel purified band, TOPO TA cloned and sequenced).

Clone 1: sequence at 5' end was the reverse complement of the 3' end - i.e. my reverse primer

Clone 2: sequence at 5' end was forward primer and 3' end was reverse primer. Seems correct, but blast results show sequence is similar to some mouse cloned product.

Clone 3: sequence at 5' end was the reverse complement of the 3' end - i.e. my reverse primer


The sequences matches the primers are 100% correct but not sure what the sequence is between the primers. The sequencing peaks look clean, sharp and good height. All sequence are the same size. Clone 1 is about 98% similar to clone 3 but both have no similarity to Clone 2. My non-template control does not produce anything.

No idea where to start, except to start over?

Sample problem - mRNA contamination?

Primer issues? Primers are reported in the literature.

Cloning or sequencing issue?

Thanks!

54 Views · 1 Replies ( Last reply by pcrman )


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