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

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

Buffer to disrupt platelet alpha granules for PF4 quantification?

Hello! I want to quantify by ELISA the secreted (from platelets poor plasma) and the non-secreted (from platelet lysate) PF4 before and after TRAP stimulation. I will use the ELISA from R&D...

03 March 2021 1,499 2 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

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 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

How optimize the Size Exclusion Chromatography?

Hello! I'll do a size exclusion chromatography, but I only have an open column, and I'll perform the peptide extraction from yeast, using buffer lysis (sodium phosphate 50 mM/NaCl 30 mM/DNAse and...

01 March 2021 2,215 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

Why are my bands weird in this Agarose gel?

Can someone please give me some possible things that could go wrong? Here is my recipe: 0.5g Agarose 50 mL of TAE 1x 1 uL ethyl bromide. Gel was run at 100V for 1 hour. The buffer used is also TAE.

01 March 2021 9,952 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

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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

What do I do with DLS peaks that are from the buffer?

I'm looking at the aggregation of my protein sample using DLS. Unfortunately, my buffer (20mM HEPES) also results in a set of peaks. These are at approximately 1 and 1000 d.nm. The lowest peak...

01 March 2021 9,015 2 View