Designing forward primer for mi cDNA?

I am using Agilent miRNA first strand synthesis kit (cat#600036) for preparing mi cDNA from total RNA. A universal primer is provided with the kit sequence of which is not given. i want to study...

07 August 2018 2,227 2 View

No signal in SYBR Green QPCR for miRNA quantification?

Hello everyone ! I am using mirVana miRNA isolation kit for the isolation of total RNA containing small RNA. Further i used Agilent first strand synthesis kit(cat#600036) for the synthesis of...

07 August 2018 750 5 View

Problem with PCR amplification?

i tried gradient PCR for a new set of primers and got the amplification. However, when i tried running the same set of primers on a single annealing temperature, it showed no or weak...

10 November 2016 9,245 14 View

Can anyone tell how to troubleshoot no amplification in PCR?

I am trying to amplify my genes of interest.Though I got the amplified product of the desired size on my first attempt , i am not able to get the amplification again. What might be the problem and...

07 August 2016 9,433 3 View

Problem with RNA isolation from Breast tissue ?

02 March 2016 1,431 3 View

Why do I get peaks in NTC in qPCR ?

I am performing real time pcr for ACTB. I am getting a ct value of 17-20 in my samples. However , i am getting a peak in NTC too with a ct value around 33-35. Also the dissociation curve for NTC...

10 November 2015 3,707 8 View

What factors should be considered in miRNA -target prediction?

06 July 2015 8,356 6 View

Can anyone help explain double bands in PCR?

06 July 2015 4,069 6 View

How can I select a good endogenous control for miRNA qPCR?

I have to perform qPCR for miRNA isolated from colorectal tumor and adjacent normal tissue samples. Is U6 snRNA a good endogenous control for qPCR?

05 June 2015 3,646 9 View

Do you know how I can process TCGA data?

I have downloaded the miRNA seq data for colon adenocarcinoma from TCGA. If I would like to see the expression of a particular miRNA across all the samples, how should we proceed?

03 April 2015 5,513 1 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 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

Is there any one who can provide a donation of HPV detection and genotyping kits in exchange collaboration fors?

Warm greetings My name is Ayichew S. and I am doing my Ph.D. in Medical Microbiology (on Human papillomavirus). At this stage, I have almost completed a clinical sample Cervical swabs)...

25 February 2021 1,660 3 View