Which forward primer I should order?

I have been trying to design primers for microRNA expression. The image shows that there are 2 sets of forward primer (Tm calculated in two different manners) and one universal "reverse primer" as...

10 November 2018 5,216 0 View

How can I characterise HMEC cells?

I have a set of breast cancer cell lines and will use HMEC as a control population. Since HMECs have the tendency to show unusual characteristics along with the passages so I would like to know...

07 August 2018 6,616 0 View

Pros and cons regarding expression profiling of microRNA-SNP using breast cancer cell lines?

I have found a SNP in a microRNA gene from my association study and now want to do the expression analysis in breast cancer cell lines (sub-type specific). What are the factors should I consider...

04 May 2018 674 0 View

How can I view .map & .ped files in command promt in Windows using PLINK?

I have got .bed .bim and .fam files from a database and need to see whats inside. But I don't know the command to do so. As well as I would like to view my own .ped and .map files in the command...

08 September 2017 8,758 0 View

In vitro assay to screen anti-arthritic property of drug compound?

09 October 2016 9,263 4 View

Which cell-cycle phase is the best for drug treatment?

I have used the drug as a broad term here. Specifically my focus is on those drugs which have antioxidant property. Is there really any time point at which cells will respond best to the...

07 August 2016 4,857 0 View

HUVEC starvation, time and with or without serum?

To get the best ROS generation (i am expecting 40-50% increase than that of control), how long my HUVEC should be starved and what would be the serum percentage? i am using EBM-2 (Lonza) without...

06 July 2016 9,227 5 View

HBSS, with or without Ca2+/Mg2+ for DCFDA assay?

06 July 2016 1,167 5 View

Why HUVEC is giving low amount of protein?

Trying to measure protein concentration for Lucigenin assay. But even culturing at more than 80% confluency, after centrifugation and sonication getting only 330ug/ml of protein. Hard to maintain...

05 June 2016 1,227 2 View

Why did TNF-alpha treated HUVEC not generating significant level of ROS ?

I am culturing HUVEC up to 80% confluency (EGM-2; Lonza), then let them serum starved (EBM-2; Lonza) for 24hr and treated with TNF-a (10ng/ml) for 30min. With DCFDA I didn't get that much of...

05 June 2016 1,975 6 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Why am I not able to use all VEP programs for the in silico analysis of avpr1b SNP's?

Good day researchers I am busy analysing the predicted effects of certain SNP's in AVPR1b. When using the VEP prediction programs (SIFT, Polyphen, FATHMM, LRT, Provean, Mutation Assessor, and...

28 February 2021 5,197 1 View

What is the difference between genotype and haplotype data in the ped format?

Hi everybody In the ped format for genotype, alleles of any SNP are represented by two columns (one for each allele, separated by a space). Is a column sufficient for the haplotype to...

27 February 2021 1,965 1 View

Can MicroRNA cross the blood brain barrier?

In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?

24 February 2021 7,064 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

FDC and FRC cell populations in mouse spleen?

I was trying to isolate fibroblastic reticular cells (FRC) and follicular dendritic cells from mice spleen, but kept getting poor yield. I found from literature that the frequency of FDC is around...

17 February 2021 3,506 4 View

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit...

17 February 2021 8,986 2 View

Why non-specific band appear on T-ARMS PCR?

Hi everyone. We want to know how can I remove the non-specific band amplification on this variant. Although the amplification of non-specific band is not as intense as the band that we want, but...

09 February 2021 588 2 View

Sequence of an unknown Oligonucleotide ?

Hi All, I do have a single-stranded oligonucleotide of 100 - 150 bases synthesized and the sequence is not known. How would I proceed to know the sequence .

04 February 2021 5,614 3 View

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...

03 February 2021 9,330 3 View