Can a sample positive for E. canis by smear test, be negative by PCR test?

We received three Dog blood samples for E. canis PCR test. The DNA was extracted using QIAamp DNA blood mini kit and sujected to PCR. The template DNA was subjected to two different PCR using...

11 December 2019 6,801 7 View

Any suggestion for low cost Real-Time PCR?

I am planning to buy a real time PCR instrument for my laboratory in Chennai, India. Budget is my primary concern and I am in a great dilemma to decide which machine might be the best choice for...

07 August 2019 8,015 3 View

Discrepency between nanodrop reading and agarose gel electrophoresis interpretation?

06 July 2019 1,294 10 View

What can be the possible reason for the signal observed in NTC during TaqMan probe based qPCR (Real Time PCR) assay?

Hello everyone, I am carrying out TaqMan probe based Real-Time PCR assay. I am able to observe signal peak for the NTC sample. However upon agarose electrophoresis I was unable to see any band...

06 July 2019 1,392 7 View

Which TaqMan dye combination is best suited for SNP genotyping?

I am planning to standardize a TaqMan duplex PCR assay for allelic discrimination. I intend to use FAM and TAMRA for one probe. Which dye and quencher would be best for the second probe that can...

03 April 2019 5,418 3 View

Why fungal DNA does not yield amplification against ITS-1 and ITS-4 primers?

I tried extracting DNA from an unknown environmental fungal isolate using Qiagen DNeasy extraction kit. However, I was unable to extract DNA in the first attempt. Then I tried increasing the...

02 March 2019 4,917 13 View

What could be the reason for non specific amplification of gDNA upon serial dilution?

The aim of my study was to evaluate the sensitivity the PCR assay for the in-house primers that I had designed. The concentration of the extracted DNA was 564.4 ng/uL with A260/A280 ratio of 1.97...

01 February 2019 185 2 View

Is it possible to differentiate plant tissues based on bisulfite sequencing methodology?

My objective is to identify the tissue of origin used for the plant extracts preparation. Is it possible to extract DNA from any provided plant extracts and subject it to bisulfite sequencing and...

01 February 2019 210 3 View

How upload a sequence with novel mutation in GenBank?

04 May 2016 4,605 6 View

How can we identify T to A mutation using High Resolution Melting analysis?

The Melt-Curve analysis are based on the GC content of the sequence. But, in case of 'T' to 'A' mutation or 'A' to 'T' mutation there is no change in GC content of the sequence. Then how can we...

06 July 2015 222 8 View

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02 March 2021 3,060 3 View

How to normalise data on OPM package from R Studio ?

Good afternoon, I recently used OmniLog from BIOLOG for my experimentations : I tested the metabolism of different strains on 2 types of plates. I have 16 strains of 3 different groups...

02 March 2021 3,584 1 View

How to grow E.coli on the low salt LB agar plate with 50 or 100ug/ml of Blasticidin?

Hello, I would like to ask how to grow E.coli which resists 50 or 100ug/ml of Blasticidin. I have tried many methods by using low salt Luria-Bertain(LB) agar plate. However, no colonies grow. Let...

01 March 2021 2,027 2 View

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28 February 2021 9,936 2 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

When is it unnecessary to change organisational culture?

As a former management practitioner, I recall a generally accepted view that changing organisational culture is difficult, so you should only attempt it if the organisational change that you are...

27 February 2021 2,595 8 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

What is the best surface to culture cells isolated from a skin biopsy?

Hello, recently I am having some difficulties with maintaining the cell culture of primary cells isolated from a skin biopsy (normal and malignant). The coating of culture plates with Matrigel (a...

26 February 2021 6,151 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

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View