How long I can storage cells after fixation with paralformaldehide (PFA)? Is okey for long term?

01 February 2017 8,979 0 View

Why I got smear in my PCR product using kod plus?

10 November 2016 6,515 5 View

How to separate 404 bp with 344 bp for dna product PCR?

08 September 2016 1,243 3 View

What the reason rasio vector : dna is 1: 90 by using vector p.BluecriptSK-II-ecoRV?

06 July 2016 950 7 View

Can anyone help with running gel for purification by gel extraction?

Dear All I have done gel extraction my PCR product (2300bp), but the final I lose all my DNA. Here my following step. Ethanol precipitation (using glycogen, Na-OCH, ethanol) My PCR product 50...

06 July 2016 4,244 4 View

Is no problem if I do Ethanol precipitation with composition glicogen, Na-asetat, before and after phosphorylation PCR product by T4 PNK ?

Dear all, Is there anyone have experience do ethanol precipitation PCR product prior to phosphorylation by T4 polynucleotide kinase? According literature ethanol precipitation by ammonium asetat...

06 July 2016 3,195 3 View

Is it possible to Clone ArfGAP protein with SH3 domains, ankyrin repearts & pleckstrin homology domains (ASAP1) in neuron brain mouse tissue?

Dear All I have plan to clone ASAP1 from neuron brain mouse tissue. Is there anyone have experience cloning ASAP1 with cDNA from neuron brain mouse? I already design primer and check by primer...

05 June 2016 216 0 View

The meaning any, self, pair any and pair End by using primer3plus ??

04 May 2016 9,290 3 View

How to solve Unexpected band after amplification PCR ?

I have done amplification by PCR for my gene, but unexpected I got several band higher expression than my band, I have try changed my condition annealing time (Tm) but the result still same,...

04 May 2016 7,280 20 View

How to check cDNA mouse embryonic fibroblast gene expression ?

Dear All Is there any software to know gene expression high or low in mouse embryonic fibroblast? Best Regards

03 April 2016 1,123 12 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

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View