TOPO or pGEM vector for rhizophere 16S library?

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Please what is the best solvent to dissolve heme?

I have tried NaOH and NH3 so far but can't dissolve well.Thanks for the assistance. I can be contact personally on this email:[email protected]

04 January 2021 5,528 2 View

PVDF membrane degradation?

Hi, I'm performing western blots on different samples and I'm facing a some problems after eletrotrasnfer with PVDF membranes. In general, during the incubations with the primary and secondary...

18 November 2020 6,412 7 View

What to do when you found a published paper that doesn't belong to you but has your name as the author?

I found a published paper online (attached here) that bears mine and colleague's names as authors but the paper doesn't belong to us. The work was claimed to be carried out in Kenya. We are in...

22 March 2020 4,860 7 View

Do you know a (validated) instrument for the assessment of state embarassment?

I've found quite a few scales for vicarious and personal embarassibility as a personality trait so far (i. e. Kelly & Jones, 1997; Maltby & Day, 2000; Uysal et al., 2014...). As I'd like...

24 February 2020 8,142 4 View

Are Peak Areas of different silylations comparable?

If I have an analyte, that has different silylations, like 2-TMS and 3-TMS, how can I compare the peak areas? Can I simply sum them up? What if theres a different ratio of 2-TMS/3-TMS in my...

03 December 2019 273 4 View

What is a good topic for Blockchain applications in IoT?

Hello everyone, at the moment I'm looking for a topic for my bachelor thesis. I was thinking about Blockchain and Smart Contracts for cyber-physical Systems, in order to use it for Smart Factory...

23 October 2019 3,694 5 View

How to carry out western blotting for a 233 kDa protein effectively?

Hi, I want to carry out western blotting for a 233 kDa protein. I was told to use a tris-acetate gel due to the size of the protein. However, I was wandering would I be able to detect the...

25 March 2019 6,308 4 View

How to set up Coupled Displacement Boundary Conditions in COMSOL?

Dear Community, I recently switched from ANSYS Mechanical to COMSOL Multiphysics. So far I was able to transfer my models to the new platform but I yet miss one feature of ANSYS in COMSOL. I hope...

29 January 2019 4,937 3 View

SpeedVac of DNA and salt concentrations: downstream effects?

I often use a speed vacuum concentrator to concentrate my DNA prior to restriction digests, DNA ligations, NGS library preparation or target captures. Usually, my DNA is eluted in 10 mM Tris-Cl...

13 November 2018 6,049 3 View

How do I calculate NDVI images with different thresholds using Sentinel-2 image?

I want to set different thresholds for calculating the NDVI images using Sentinel-2 for the purposes of fishing out river sediments in the Dry and Wet seasons. I am using the Semi-automatic...

05 November 2018 694 2 View

Would fecal samples stored at -20C for over a year impact metagenomic analysis?

Dear All, I have access to some rodent fecal samples that have been stored at -20C instead of -80C. Anyone have experience with the impact of stowage condition on comparative metagenomic analysis...

01 March 2021 2,512 1 View

How do I check the quality of my viral RNA prior to metagenomic sequencing?

I am doing metagenomic sequencing without enrichment for an RNA virus. After my RNA is extracted from the blood I am doing library prep with the NEBNext Ultra directional RNA library prep kit for...

16 February 2021 5,101 2 View

Does DNA A-tailing survive after agarose gel electrophoresis and purification?

Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...

04 February 2021 6,051 8 View

How can I solve this problem with agarose gel DNA purification of bigger (1-4kb) fragments ?

Hi everyone, I perform a PCR + Restriction Digestion (single cut, same at both sides) over a metagenomic library. After that, I am trying to purifiy DNA fragments between 1-4kbp long from the...

24 January 2021 2,190 5 View

Isolation protocol for rice rhizosphere microbes?

I want to extract various microorganisms from the rice rhizosphere, particularly bacteria and fungi. Can anybody recommend an isolation protocol specific to the said purpose?

19 December 2020 3,541 3 View

Could anybody provide insights the best tools to create a custom database for taxonomy assignment of metagenomic data?

I am interested in the different tools that can be used to create custom databases for targeted sequencing and how to trim the databases based on the amplicon size? Also, should custom databases...

02 December 2020 3,105 3 View

Does Shannon index represent largest diversity index when the individual belongs to different species and vice versa?

The shannon wiener index and Simpson indices include the measurement of community heterogeneity. Shannon wiener index bapplied information theory. The main measuring object of information theory...

22 October 2020 1,053 7 View

What is the best technique to identify native bacteria groups present in soil?

I am lay in this subject. Is DNA sequencing the best way to identify the different types of soil bacteria present in soil samples? Ideally, I'd like to find a technique that allows me to identify...

29 September 2020 4,609 9 View

Is the first clean up necessary in Illumina MiSeq Library Preparation?

I'm trying to streamline my metagenomics protocols for my lab. I have previously followed the 16S protocol...

16 September 2020 5,234 6 View

How to use Vegan::capscale to calculate and plot distance based- redundancy analysis (db-RDA) and add correlation cut-offs in the results?

I have a dataset with relative abundances of Genomes against their COG-ids. It is a large data frame with 4214 rows (COG-ids) and 1313 columns...

13 September 2020 2,334 1 View

Matthew G. Bakker

You might consider using a high-throughput sequencing method rather than constructing a clone library at all. Or do you need the longer sequence reads for some reason?

Felix Moronta-Barrios

You're right! I will contact to the company which sequence my samples (Macrogen) and ask them advice and budget. (hope this were not stratospheric). Tks.