Could anyone help me with PCR to Mycoplasma detection?

More Bruna Rodrigues's questions See All

Physical-chemical formation of stalactites/stalagmites. what promotes the formation/growth of these structures?

I know, or think I do, the basic idea behind these natural structures, as from what I know they form due to water dripping from the top leaving a salt deposit behind. But started to wonder about...

22 February 2021 1,400 4 View

Can the intensity of this broad band in XRD mask other crystaline phases present?

I obtained a XRD diffractogram with significant broad band at low 2theta levels, also around 30 2theta another smaller broad band appears. It all points out to the existence of an amorphous phase....

19 February 2021 8,414 4 View

How to measure a Neural Network complexity?

Does anyone know how to measure the complexity of an artificial neural network? Could you provide me references? Thanks a lot!

25 January 2021 2,710 10 View

What is the best method for removing power-line interference (PLI)?

In the paper A wavelet-based method for power-line interference removal i... we propose a new method for removing PLI. This method consists in identifying the ECG and noise frequency range for...

29 December 2020 9,233 3 View

Tools for analysis of interaction between lncRNAs and proteins/miRNAs?

Hi! I would like to know what tools I could use to analyze interactions between a new lncRNA and possible protein/miRNA targets. The biggest problem I had to find such tools is that the RNA we...

20 November 2020 8,331 3 View

In which dry organic solvent/solvent mixture molten 1H-imidazole could be precipitated at 100%?

I am trying to isolate an allkylated imidazole from a mixture containing unreacted 1H-imidazole. The problem is that imidazole is slightly soluble in my targed molecule. Furthermore, I need to...

04 November 2020 4,513 3 View

Should I exclude a paper classified as low quality by the Newcastle-Ottawa tool from a systematic review without meta-analysis?

Hello! I'm working on a systematic review (without meta-analysis) and when evaluating the quality of the papers only one presented low quality. I did not find guidelines in the instrument itself...

04 November 2020 6,564 7 View

When will we have access to Brazilian agricultural censuses microdata publically available? Can Google and Microsoft help it?

Microdata are a wonderful way of analyzing specific issues in an heterogeneous environment. Brazil is wonderfully diverse country, and its countryside is also extremely heterogeneous. We have...

25 October 2020 8,900 2 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 it possible to use a muffle furnace without N2 atmosphere to make Biochars?

Greetings Everyone. I am reading several articles about biochars and notice several use N2 atmosphere inside the muffle furnace. can you produce biochars without this N2 atmosphere or other inert...

17 August 2020 8,992 6 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

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 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

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

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 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

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Houda Kawas

Dear Bruna Rodrigues

Pls. find the attached files, may help in your research, good luck

http://www.ctcusp.org/pdf/inctc_posters/30.pdf

http://genome.cshlp.org/content/4/4/199.full.pdf

http://link.springer.com/article/10.1007/BF00127903

http://icmr.nic.in/ijmr/2013/august/0811.pdf

https://www.researchgate.net/profile/Jens_Boman/publication/11880269_Dowell_S_F_et_al_Standardizing_Chlamydia_pneumoniae_assays_recommendations_from_the_Centers_for_Disease_Control_and_Prevention_USA_and_the_Laboratory_Centre_for_Disease_Control_Canada_Clin_Infect_Dis_/links/56546f1508ae1ef92976a1a1.pdf

http://bcrj.org.br/wp-content/uploads/2015/08/Clique-aqui.pdf

https://www.researchgate.net/profile/Martin_Altwegg/publication/10875508_Development_of_a_multiplex_real-time_quantitative_PCR_assay_to_detect_Chlamydia_pneumoniae_Legionella_pneumophila_and_Mycoplasma_pneumoniae_in_respiratory_tract_secretions/links/0fcfd50bf3e45944cb000000.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3949616/

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3463982/pdf/10616_2004_Article_5113481.pdf

Stephen Gillespie, Peter M. Hawkey. 2006. Principles and Practice of Clinical Bacteriology John Wiley & Sons,Pp. 620. ISBN 0470035323, 9780470035320

R. Ian Freshney. 2011. Culture of Animal Cells: A Manual of Basic Technique and Specialized ApplicationsWiley Online Library: Books , Pp. 768 . ISBN 0470649356, 9780470649350

Regards

Prof. Houda Kawas

Article Standardizing Chlamydia pneumoniae Assays: Recommendations f...

Article Development of a multiplex real-time quantitative PCR assay ...

Ram Gopal Nitharwal

Hello Bruna,

Try the following.

1) use a gradient PCR to know the exact conditions for pro,er annealing.

2) Lot of smear is there in the PCR. Titrate the magnesium concentration.

3) Is mycoplasma GC rich or AT rich? keep this in mind for choosing the strategy.

4) Try to use Fusion DNA polymerase. This is faster and more specific them Taq.

Hope it works for you.

Cheers!

Bharathy Sukumar

Try to standardize your annealing temperature by using gradient PCR.