Why is rbcL not getting amplified in macroalgae?

More Satej Bhushan's questions See All

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I have tried to isolate DNA from Padina but it is not getting amplified using markers that are working fine for other algal samples. Is there any specific method or precaution which should be...

31 December 2013 182 4 View

Smearing in agarose gel of PCR product?

How can we explain smears in agarose gel when PCR products are run? It could be mistaken for DNA.

31 December 2013 1,594 39 View

Which technique can identify gene location?

How to identify whether a particular bacterial gene is chromosomal or on plasmid, before sequencing?

10 November 2013 323 6 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

How to increase a model accuracy ?

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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?

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Which cell line should we use to perform biological dosimetry experiments?

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How to address this issue in the analysis of Real-Time PCR results?

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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

Any research papers for extraction of nutrient deprivation on macroalgae? Analysed by HPLC-UV and GC/MS. Possibly from macroalgae genus Rhizoclonium?

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28 February 2021 6,477 1 View

MiRNA qpcr problem?

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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...

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Sunil Kumar Sahu

Which primer you are using for rbcl? If you can provide more details regarding PCR program and concentration of DNA then some suggestions can be made Satej.

Satej Bhushan

94C 5 min, 94C 1 min, annealing at 50C for 1.5 min, & then 72C for 2 min (35 cycles) and final extension for 7 min

concentration varies between 50-100 ng/ul

Beivy Kolondam

Hi Satej, how did you extract the DNA? How long did you do cell lysis? Were you using proteinase K? Sometimes you have lots of nuclear DNA but no chloroplast DNA. You must incubate longer time in cell lysis solution and proteinase K (approx. 1-3 hours in 55'C).

@Beivy

Hi, I used algal DNA extraction kit. I used proteinase K as well but followed manufacturer's protocol which was 30 min for lysis. Thanks, i will try with increased time.