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

TaqMan SNP genotyping: Is there a need to use the UNG activation step?

I am using TaqMan probes for SNP genotyping on LightCycler 480. I am facing some difficulties in resolution of heterozygous genotypes. The software can not identify it as " both alleles"...

10 November 2013 6,706 2 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

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

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

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View