How to make quantification of specific bacteria added to the soil?

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Is it Ok to decline to write a chapter for a IntechOpen to the lack of experience?

I have been asked by IntechOpen to write the chapter for a book they are going to publish, but as I don't have enough experience in the area they proposed me to write I declined their offer. Now...

24 February 2021 2,802 4 View

How to inerpret whether PCR went OK?

Dear friends! I am back to work after 3 years career break. Now performing a PCR, feeling like a newbie... Please, help to understand whether RT-PCR reaction "worked" or not. Which results should...

14 February 2021 3,129 8 View

Which fungicide are good for primary cells? Does it yeast contamination? How to get rid of them without killing the cells? How to get rid of spores?

Hello! Our lab is working with primary cells which we are isolating from the colorectal tissue. During the isolation, we are washing the tissue with pen-strep and then also add some pen-strep in...

07 February 2021 8,258 3 View

EMOJI-POLL-3 (Emoji-Association): Can you join the survey?

https://docs.google.com/forms/d/e/1FAIpQLSfaaCWZZqMIhoz5yj_5dxggvb6-cISx3GUHssYUWZWKbpx3UA/viewform Dear colleagues! We ask you to participate in a series of surveys related to the scientific...

26 January 2021 5,969 3 View

The same lC-MS for Forensic Toxicological and Chemical expertises?

Dear Colleagues! How correct and appropriate is it to carry out a forensic toxicological examination of drugs and psychotropic substances in biological objects (in blood, urine, hair) and a...

24 January 2021 4,489 2 View

GC-Headspace (HS-GC) chromatography for Forensic toxicological and Food expertises?

Dear Colleagues! How correct and appropriate is it to conduct forensic toxicological and food examination of volatile substances (ethanol, methanol, propanol, ethyl glycol, etc.) in blood and...

24 January 2021 5,645 2 View

How to choose mouse cell line to work on?

Hey, I am looking to start working on mouse cell line for some genomics experiments. I have the following requirements: 1) Normal chromosomes composition. 2) Possible to transfect. 3) Possible to...

03 January 2021 4,387 2 View

How to describe the method described below?

I do research on the relationships between universities and local governments by asking the research question (how) can universities contribute to the design (and implementation) of public...

29 December 2020 4,522 12 View

How can I change the media for THP-1 cells without dislodging them from the surface of my well plate?

I am working with THP-1 cells at the moment and am having trouble with maintaining a high confluence once I seed them on a 24-well plate. I usually try to seed around 100,000 cells per well and...

29 December 2020 1,567 3 View

What glass vials do you use for small volumes (400ul-2000ul) that fit in the centrifuge?

I am planning some NMR experiments that require glass vials (due to chloroform). I am looking to find glass vials that would take 400-2000ul volumes, fit in the centrifuge and allow me to separate...

01 December 2020 3,259 2 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 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

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

How to accurately quantify chitin?

Hi all, I have been working on the quantification of chitin in insect samples and feed for a while but I can notice that the quantification methods (e.g. exposure to HCl, NaOH, and weighing sample...

01 March 2021 1,843 2 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 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

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Walid Hassene Hamri

Hi Anna,

You'll find below some articles that may help you.

https://www.nature.com/articles/s41598-017-05260-w

Article Optimization of a Method To Quantify Soil Bacterial Abundanc...

https://www.jove.com/v/10099/culturing-and-enumerating-bacteria-from-soil-samples

Chapter Methods to detect and quantify bacteria in soil

Frederic Lepretre

Hi Anna

I'm sure you'll find in this poste the "how to"...

Article Detection of bacteria in soil by direct DNA extraction and PCR

designed primers can afterwards be tested by in silico PCR at NCBI site.

fred

Abhijeet Singh

Anna Marzec - Grządziel

You can also try to search some articles or look into your references for the primer pair. There might be some some species specific primers already designed which can serve your purpose. If not the primer itself, you will surely come to know about how to design the species specific primers for "qPCR" or normal PCR.