We have encountered a recurring issue with sequencing. Has anyone experienced something similar?

22 February 2022 1 5K Report

Regularly on the plasmid matrix, the following sequencegrams began to be obtained. In your experience, what could it be? Thanks!

Katie A S Burnette

Looks to me like you didn't start with enough PCR product. Add more and try again.

Badges
Science method

More Sergey Rykov's questions See All

How do we pick data for determination of Validation Acceptance Criteria?

Hello, colleagues! There is commenting open for new upcoming edition of USP 1033. Validation target acceptance criteria is now different from what it used to be and it doesn't include Cpm....

23 July 2024 7,292 3 View

Hydride zirconium properties?

Could you present the review or nowadays status state investigation of thermophysical and physico-mechanical properties of hydride zirconium (ZrH2)? Where can I find out properties data tables?

02 April 2024 2,025 0 View

Ansys APDL_Receiving resulting heat flow in finite element?

Could anyone explain *how to* or give an example of receiving heat flow (watt) for the FE element? I understand that *GET command is useful like *GET, PAR, ELEM,NO_ELEM,...,SMICS but can't get...

01 April 2024 6,562 2 View

Is it possible an unspecific binding of streptavidine or peroxidase?

I have a question for the professionals. The essence of the problem: I fix the oligos on a plastic substrate, they play role of primers in solid-phase PCR. I carry out a one-step PCR with...

28 March 2024 9,110 2 View

Simulation of temperature field in a brick, taking into account thermal expansion with ANSYS Mechanical APDL? (Thermomechanics in details)?

Problem to model a simple (at first glance) stationary problem of thermomechanics in an elastic formulation in ANSYS Mechanical APDL. There is a simple block (brick), with spatial non-uniform...

28 March 2024 7,709 2 View

Ansys Mechanical APDL__4D table input (axes and indices)?

Hello colleagues! Several days ago I used to struggle with tech problem of 4d table input QV = QV(X,Y,Z,TIME) using Mechanical APDL commands script. I haven't solved it yet. 1) Standard technique...

22 March 2024 3,515 0 View

I am looking for B-H curves of GO steel M080-23DR (Domain Refined)?

I am looking for B-H curves of GO steel M080-23DR (Domain Refined). It is customary to think that the "technical saturation" of GO steels is near 2.0 – 2.04 T. However, there are some indications...

04 March 2024 7,393 0 View

How to get solution for house-selling task (or task of selling an asset) with conditions “finite horizon, recall is allowed” ?

House-selling is one of the typical tasks of the Optimal Stopping problems. Offers come in daily for an asset, such as a house, that you wish to sell. Let Xi denote the amount of the offer...

03 February 2024 4,482 4 View

What dye do you use for RNA electrophoresis?

Colleagues, tell me which dye is optimal for staining RNA during gel electrophoresis? Which one do you use in your laboratory?

21 January 2024 6,373 4 View

Are you in Public Health and interested the project: "Central Asia Alliance for Climate-Health Research"?

I seek coauthors and colleagues from Uzbekistan, Kazakstan, or other CA countries in "climate-air-pollutant-health" research. It would be great to collaborate with individuals who share similar...

28 November 2023 8,493 0 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

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

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 View

Should the amount of DNA input used for ChIP-seq library preparation be matched between the control and experimental groups?

Hi all. As a beginner in ChIP-seq experiments, I hope you understand that the following questions might be somewhat basic. I am planning to perform ChIP-seq or MeDIP-seq analysis to investigate...

28 July 2024 6,938 1 View

If my gene of interest has high GC content can it be problematic in sequencing? What kind of error is expected with GC rich gene sequences??

Gene sequencing related trouble shooting

25 July 2024 4,149 2 View

Does post-translational protein modification cause devisions on observed pI verses calculated pI?

In running two-dimensional gel electrophoresis on bacterial protein, some spots that appear to match a protein sequence have a significantly more acidic isoelectric point than the calculated pI....

24 July 2024 8,076 3 View

Are there always been barcodes, apapters and primer sequences in the FASTQ files of NGS?

Hello researchers, Sorry for my stupid question. I am learning the QIIME2 workflow for analyzing some 16s amplicon NGS fastq data. I found a very nice paper with data and code public available...

20 July 2024 5,404 2 View

Promotor observation in region annotation of RNA RIP-seq?

Hello all, I extracted RNA from my samples and performed RIP-seq. After annotating the genomic regions using R, I obtained promoters, exons, introns, and UTRs. Given that my samples consist of...

18 July 2024 1,579 2 View

Why does it show two bands when I add single hairpin structures to a gel?

Hi everyone, I am working on a hybridization chain reaction (HCR) and trying to visualize it with gel electrophoresis. The visualization of the HCR works but for the single components of this...

07 July 2024 9,413 4 View

Are DNA samples with 260/280 ratio greater than 2.1 good for sequencing?

I have DNA samples (extracted by using Qiagen kiit), the 260/280 ratio is consistence 2.1.. for most of the samples. Can I proceed for the whole genome sequencing with these values? NGS.

07 July 2024 3,751 4 View