Looking for complete sequence of pSRK plasmid ?

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How to quantify two different fluorescent tagged bacteria (Sm1021)from root nodules of Medicago trancatula in a competition experiment?

I have tagged Sm1021 with GFP and mCherry. GFP is tagged with gene. mCheery is the control and vice versa. GFP-gene, mCheery only mCherry-gene,GFP only I have infected Medicago with the above...

11 December 2018 5,164 0 View

No bands after restriction digestion purification of a low copy no plasmid ?

I am trying to clone few genes within a very low copy no (7.4Kb) plasmid. The yield is pretty low after Mimiprep from 5ml of over night culture, eluted in 30 microliter of water/TE buffer. And...

04 May 2018 7,118 2 View

How to determine ploidy by flow cytometry?

I am trying to determine the ploidy level of bacteriods (Sinorhizobium meliloti from the root nodules) taking the free-living bacteria as the reference. I would like to know how to determine the...

04 May 2017 3,478 2 View

How to design His tag primer?

I am planning to design primer for His tag. The general format of the forward primer : CGC: Restriction site (GGTACC) :Spacer(GGAGGTACTGA): Start codon (ATG): Six Histidine (CACCATCACCATCACCAT):...

09 October 2016 5,318 10 View

What is the effect of intravector multiple insertion in plant transformation vector?

Plant transformation and biotechnology

10 November 2015 3,729 1 View

Can anybody help me to analyse LC MS MS data in scaffold file ?

08 September 2015 478 0 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

Can formaldehyd-fixed gelatin be melt?

I am puzzled about the properties of gelatin, when used for tissue-embedding. Hard gelatin can be melt again by temperatures about 40°C. Formalin-fixed gelatin is like crosslinked protein and...

07 August 2024 1,686 1 View

How to fix errors in my heat transfer steel structure with reinforced concrete slab model Abaqus?

I have modelled a steel structure using beam elements in Abaqus and attached to this structure reinforced concrete slab. The analysis that I am making is heat transfer of the structure. The...

07 August 2024 1,028 0 View

Unusual intensity drop in some sections of chromatograms in DDA?

Hi, we have measured tryptic peptides using both DDA and DIA method on QExactive. In DDA replicates i saw unusual intensity drops occurring at the same sections of chromatograms in DDA replicates...

07 August 2024 3,218 4 View

Microtome tissue sectioning - How to prevent chatter of paraffin tissue specimen sections?

Hello everyone, I have recently started using the microtome device for sectioning of paraffin-embedded mice lung. While I had some success in sectioning and observing proper ribbons, some of my...

06 August 2024 666 4 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

What should I do with parameters that are not relate to my simulation in MyLake model?

I want to Estimate surface heat fluxes using MyLake, but I don't have all the initial values in model parameters section and other sections,is there a way?

04 August 2024 1,537 1 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View