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 design vector with miRNA variant (SNP) for expression analysis?

I have found a miR-SNP which is significantly associated with my breast cancer case-control cohort. Now I want to see its expression in different breast cancer cell lines to find out the best...

03 April 2018 5,145 4 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

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 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

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 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

Genescan gives 3 peaks instead of one?

Hi all, I am doing a genescan analysis for a deleted exon in the patient sample. The amplified region is 215 bp. In the patient sample because of homozygous deletion, no peak is observed while in...

23 February 2021 5,645 1 View