How to prepare NALC buffer for liquefaction of non-TB target sputum?

Can anyone tell me if NALC-NaOH is suitable for liquefaction of sputum for real-time pcr amplification of non TB pathogenic bacteria and virus. Otherwise how to prepare NALC-NaOH or NALC PBS...

31 December 2018 2,649 0 View

Ho can I DNA extraction from tallow or beef fat??

Hi.. I need a suggestion to isolate animal DNA from tallow or beef fat.. Proceesed tallow doesnt allow recovery of DNA as it has been boiled over 110 deg celcius. Still i need a method to work...

11 December 2015 5,465 3 View

Two or three step annealing for low Tm probes in AT rich region?

I want to do to genotyping of the a sample and designed the probes with many softwares but all are giving me probes with tm below 59 deg. as per qPCR guidelines I tried making primers at 4-5 deg...

06 July 2014 5,365 1 View

Any suggestions for genetic risk prediction?

I am trying to understand the implication of some SNPs responsible for disease in cattle. I want to calculate and predict the risk of disease from risk allele of an SNP in an individual animal...

10 November 2013 7,154 1 View

Do I need to activate UNG in Taqman Universal Master mix?

I am doing SNP genotyping on Lightcycler 480 using taqman probes and Taqman Universal Master Mix with UNG. Sometime the software get a low signal for heterozygous samples and the software is not...

10 November 2013 8,833 5 View

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05 June 2013 4,084 1 View

TB detection kit for Roche Light Cycler

Can anyone please suggest a kit for Mycobacterium tuberculosis detection on Roche Light Cycler 480 II?

03 April 2013 5,084 1 View

Blood parasite detection using PCR?

Can anyone tell me how long it is good to store animal blood without reducing the parasite detecting efficiency? I tried isolating whole blood DNA after 2 months of storage and it is not giving...

01 February 2013 2,438 5 View

Does anyone know a way to remove genomic DNA contamination from Mitochondrial pellet using DNAse treatment?

I am facing the problem of genomic DNA contamination in my mitochondrial DNA isolation from insect tissue. I want to include a DNAse treatment of my whole mitochondrial pellet before proceeding...

11 December 2012 5,826 7 View

Can anyone working on IonTorrent suggest a good way to reduce polyclonal ISPs?

I have run some chips (318) on Ion Torrent but I am getting high polyclonal beads many times. I have tried reducing my library concentration but it has not worked for me, can anybody suggest a...

10 November 2012 7,386 7 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 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

What solvent can I used for Paal Knorr reaction between 6-amino hexanoic acid and 2.5 hexanedione?

I am try to make the Paal Knorr reaction between 2.5 hexnedione and 6-amino hexanoic acid, my problem is type of solvent to use, because 6-hexaminohexanoic acid is soluble in water but I am not...

02 March 2021 4,443 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

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

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

01 March 2021 5,311 1 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

I want to performe MALDI-TOF MS for bacterial identification, I would appreciate it if anyone can tell me the matrix solution preparation protocol ?

based on what I've studied on multiple papers, for peptide and protein identification Alpha-cyano-4-hydroxycinnamic acid (CHCA) is an appropriate matrix to use, I also know that the CHCA powder...

26 February 2021 6,799 3 View