Small RNA Next Generation Sequencing?

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How to identify conserved miRNA if related data is not available in miRBase?

I have miRNA data of a biotrophic fungal pathogen, I would like to identify conserved miRNA by homology approach, but in miRBase the fungal miRNA data is not available. Kindly provide your...

01 February 2019 5,903 2 View

How to amplify fragment of approximately 2500 bp?

Hi I want to amplify the fragment of 2500 bp, but everytime it failed. I performed touchdown PCR and I got amplicon size of 500 bp. What might be the reason that I am unable to get the desired...

08 September 2016 4,586 7 View

What does number indicates at nodes in phylogenetic analysis?

Hi I recently started working on phylogenetic analysis. I made a tree the picture is attached below. What does number indicates at nodes of the tree. What more inference can I draw from the...

02 March 2016 9,329 13 View

Transporter prediction based on amino acid sequence?

I have an amino acid sequence of 110 which codes for a functional transporter. Utilizing which online tool and what are the possible way by which I can confirm the structure, functioning, or any...

01 February 2016 1,057 3 View

Why there is difference in band position in uncut plasmid and restriction digested?

I ran uncut plasmid with my insert and digested, what will be the reason for the difference as the uncut plasmid consist of the insert and the digested plasmid released the insert, so the band...

31 December 2015 5,985 5 View

Double bands after restriction digestion?

I got 2 bands after restriction digestion of unknown gene fragment, does it mean that my gene has restriction site apart from vector. Can I assume that addition of these two band will give me the...

31 December 2015 9,899 5 View

Is there any need to use a single digested plasmid for a good gel image?

When we use the plasmid with insert, uncut plasmid, double digested plasmid, and ladder for gel pic, why there is a need to use single digested plasmid in making a good gel image.

11 December 2015 1,668 3 View

Can someone help with blue-white screening?

I have been doing TA cloning for quite sometime. My sample is giving me only few white colonies. I took two control, the first control is plasmid with insert, which resulted in full plate white...

07 August 2015 3,483 4 View

Is there a problem with my RNA pellet?

Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...

11 August 2024 7,082 3 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

Strugglling with m6A dot blot any suugesstion ?

I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...

10 August 2024 8,539 5 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

RNA Extraction Using Hot Borate Method No Longer Working?

I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...

08 August 2024 9,882 2 View

How can I apply boundary conditions in an orthotropic steel deck numerical model using ABAQUS software?

I am trying to simulate vehicular loading on an orthotopic steel deck bridge section in ABAQUS software. The red arrow mark in the attached figure indicates the direction in which the vehicle will...

08 August 2024 719 0 View

Does Anyone have expertise in in vitro transcription and RNA pull down assay?

I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.

07 August 2024 6,622 1 View

Is there anyone with experience in TEM analysis who can assist with a manuscript for an upcoming journal?

Hello dear colleagues, We have prepared a manuscript on NiTi-based alloys and are seeking a second opinion on our current TEM results. If you are a Ph.D. holder with experience in TEM and have...

07 August 2024 9,563 0 View

Unusual intensity drop in some sections of chromatograms in DDA?

Hi, we have measured tryptic peptides using both DDA and DIA method on QExactive. In DDA replicates i saw unusual intensity drops occurring at the same sections of chromatograms in DDA replicates...

07 August 2024 3,218 4 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View