Journal: bioRxiv

Article Title: A genetically-encoded three-colour stress biosensor reveals multimodal response at single cell level and spatiotemporal dynamics of biofilms

doi: 10.1101/2022.09.23.509207

Figure Lengend Snippet: (a) Multimodal response of RGB-S E. coli to stress inducers depicted as fold changes (FC) of fluorescence signal relative to non-stressed controls. Error bars are the standard deviation (SD) of six biological replicates. ( * ) test and control samples show statistically significant difference (p≤0.05). Cultures containing the reporter plasmid were incubated for 5 h in Luria-Bertani medium supplemented with kanamycin (LB+kan), except for starvation that was conducted over 9 days. (b) Analysis of heterogenic response on the single-cell level to 2-propanol 2% (v/v) determined by microscopy analysis after 5 h incubation. Scale bar is 25 μm. (c) Representative images (obtained from b ) showing cells that display single, multimodal or no stress response. Scale bare is 5 μm.

Article Snippet: Chemicals used in this study are: EtOH 96% ANALAR, 1-BuOH ANALAR and 2-propanol HPLC grade from VWR international GmbH.

Techniques: Fluorescence, Standard Deviation, Plasmid Preparation, Incubation, Microscopy

Journal: bioRxiv

Article Title: A genetically-encoded three-colour stress biosensor reveals multimodal response at single cell level and spatiotemporal dynamics of biofilms

doi: 10.1101/2022.09.23.509207

Figure Lengend Snippet: RGB-S E. coli cells were treated with 2-propanol 2% (v/v) and sorted using FACS. Populations were sorted based on individual cell responses into four subpopulations, mono GFP, mono RFP, dual RFP-GFP, and triple RFP-GFP-BFP, with numbers (1, 2) indicating replicates. Differentially transcribed genes involved in SOS (a) , RpoS (b) , and RpoH (c) stress response pathways were analysed within the four sorted subpopulations. (d) Analysis of differentially transcribed metabolic pathways as annotated in the KEGG database of the four sorted subpopulations.

Article Snippet: Chemicals used in this study are: EtOH 96% ANALAR, 1-BuOH ANALAR and 2-propanol HPLC grade from VWR international GmbH.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

doi: 10.3390/ijms21228741

Figure Lengend Snippet: Initial testing of the tobacco rattle virus (TRV)-LAMP method. The NEB WarmStart qRT-LAMP method was tested with HPLC-purified primer mix 1 (PM1). Template = RNA purified from TRV-infected (positive, green) and uninfected (negative, blue) tuber tissues. The total reaction volume was 10 µL (including 0.5 µL template, 0.5 µL primer mix and 0.1 µL 50× LAMP dye). Positive control (PC, orange) = German Collection of Microorganisms and Cell Cultures (DSMZ) virus isolate TRV PV-0352. Negative non-template control (NC, gray) = milliQ water. For visual clarity, the amplification plots ( a ) show the increase in fluorescence with the number of cycles, and the bar chart ( b ) shows the overall Ct values. RT-LAMP tests were monitored for 1 h (1 min/cycle).

Article Snippet: Filtered supernatant was mixed with an equal volume of Rotisolv HPLC gradient grade ethanol (Carl Roth, Karlsruhe, Germany) before loading onto the columns.

Techniques: Virus, Purification, Infection, Positive Control, Control, Amplification, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

doi: 10.3390/ijms21228741

Figure Lengend Snippet: TRV LAMP primer mix comparison. NEB WarmStart qRT-LAMP tests with ( a ) HPLC-purified primer mix 1 (TRV-PM1) or ( b ) TRV-PM2 with new forward internal primer (FIP) and backward internal primer (BIP). Template = RNA purified from TRV-infected (positive, green) and uninfected (negative, blue) tuber tissues. We used 1 µL (1:10 dilution) of template together with positive and negative controls in a total volume of 10 µL. Positive control (PC, orange) = DSMZ virus isolate TRV PV-0352. Negative non-template control (NC, gray) = milliQ water. RT-LAMP tests were monitored for 1 h (1 min/cycle).

Article Snippet: Filtered supernatant was mixed with an equal volume of Rotisolv HPLC gradient grade ethanol (Carl Roth, Karlsruhe, Germany) before loading onto the columns.

Techniques: Comparison, Purification, Infection, Positive Control, Virus, Control

Journal: International Journal of Molecular Sciences

Article Title: Fast, Precise, and Reliable Multiplex Detection of Potato Viruses by Loop-Mediated Isothermal Amplification

doi: 10.3390/ijms21228741

Figure Lengend Snippet: Optimization of the potato virus X (PVX) RT-LAMP protocol. We used qRT-LAMP to amplify PVX-positive controls (PV-0014, PV-0020, PV-0847, and PV-1101), negative control (NC-0017), and negative potato tuber RNA (N11 and N12) in two dilutions (1/200 and 1/1600). We used PVX primer mix 4 (PVX-PM4) standard desalted, with HPLC-purified FIP/BIP, or with and additional loop primer. We added 450 mM betaine or 7.5% ( v / v ) DMSO to the reaction mix. 1 For the Ct value, one cycle = 1 min.

Article Snippet: Filtered supernatant was mixed with an equal volume of Rotisolv HPLC gradient grade ethanol (Carl Roth, Karlsruhe, Germany) before loading onto the columns.

Techniques: Virus, Negative Control