Anchored PCR (A-PCR) for genome walking?

i am trying to identifies unknown sequence in nitrate reductase gene from chlorella vulgaris using anchored pcr. first i extracted the genomic DNA and i did single primer extension using 0.5 ug of...

05 June 2018 8,739 11 View

Overlapping pcr of 4 fragments?

hello i am trying to overlap four fragment through use of overlapping pcr: the fragments are CMV promoter (595bp), Kanamycine resistance gene (795bp), IRES (206), and EGFP (720bp). i designed the...

01 February 2017 9,201 5 View

Live modified organism?

hello i was reading this paper http://www.cell.com/trends/biotechnology/abstract/S0167-7799(15)00088-8 and they used the cas9 protein and guide RNA without use of plasmid. is it possible to call...

10 November 2016 661 13 View

Protein Electrophoresis and Blotting for RNA page electrophoresis?

06 July 2016 1,983 3 View

What happen to microrna after inhibiting translation of mRNA?

hello everyone i am curious about the fate of microrna after inhibiting the translation of mRNA can someone provide me with information about it thanks in advance.

03 April 2016 471 0 View

Extract 3'utr from annotation file?

hello i am looking to extract the 3'utr sequences of fathead minnow fish using the annotation file and the genome. http://www.ncbi.nlm.nih.gov/genome/?term=fathead%20minnow can someone help me or...

02 March 2016 6,470 0 View

Construction of artificial microrna vector to target a gene?

31 December 2015 2,661 4 View

MicroRNA blast against miRbase?

Hello everyone I did an experiment using microRNA NGS and because of lack of funds we did the alignment of microRNA against zebarfish microrna and I got some novel microRNA. Now I want to...

11 December 2015 9,701 6 View

Does anyone know about the inhibition of microRNA?

hello I wanna ask about inhibition of microRNA expression using vector expressing microRNA sponge. i was looking for some reference about this subject and couldn't find. can someone help me?...

10 November 2015 6,587 5 View

For MiRNA knockout, should I use a bulged microRNA sponge or a perfectly complementary one?

hello everyone i wanna knockout a specific microRNA and i would use microRNA sponge and through my readings i found some researchers used bulged sponge and others used perfect sponge complementary...

10 November 2015 5,602 0 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

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 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

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