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Characterization of sporulation dynamics of Pseudoclostridium thermosuccinogenes using flow cytometry

Single-cell analysis of microbial population heterogeneity is a fast growing research area in microbiology due to its potential to identify and quantify the impact of subpopulations on microbial performance in, for example, industrial biotechnology, environmental biology, and pathogenesis. Although several tools have been developed, determination of population heterogenity in anaerobic bacteria, especially spore-forming clostridia species has been amply studied.

hAlbumin FRET 50

AH1005-50 1 Kit
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hAlbumin TRF 50

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Description: Extended warranty on a BB50-DX Bullet Blender, from 2 to 4 years. Includes priority on repairs, parts, labor and domestic shipping.

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KE785-50 50 preps
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hInsulin FRET 50

IH1017-50 1 Kit
EUR 266.00

hInsulin TRF 50

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EUR 288.00

mIgG TRF 50

MIGTR1020-50 1 Kit
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rIgG FRET 50

RI1024-50 1 Kit
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TruStrip Sample Transfer Strips, 50-ul, 50/Pk (without sample tracking dye)

STS-50-50 1 Pk Ask for price

TruStrip Sample Transfer Strips, 50-ul, 50/Pk (with sample tracking dye)

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AR02-P0011-50 50ug
EUR 727.00

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Rabies(Rab-50) Antibody

BNUM0943-50 50uL
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Description: Primary antibody against Rabies(Rab-50), 1mg/mL

Adrenomedullin (ADM/AM, 1-50), rat

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AR02-P0003-50 50ug
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Rv1009 (RpfB)-50

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Rv2450c (RpfE)-50

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Rv3875 (esxA)-50

AR02-P0008-50 50ug
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Rv2031c(HspX)-50

AR02-P0009-50 50ug
EUR 727.00

Rv3131 (NfnB)-50

AR02-P0010-50 50ug
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ERLENMEYER FLASK, 50 ML, PP

4985P-50 6/pk
EUR 50.00
Description: Reusable Plastics; Reusable Erl Flasks

ERLENMEYER FLASK, 50 ML, PMP

4990P-50 6/pk
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Description: Reusable Plastics; Reusable Erl Flasks

Rv2769c (PE27 protein)-50

AR02-P0001-50 50ug
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TR-FRET Bridge-It SAM 50

TRF-1-1-1003A 1 Kit
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slingshot homolog 1 (SSH1)(MBP-his tagged)-50

AR09-P0023-50 50ug
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DNA Gel Extraction Kit - 50 Columns

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EUR 155.00
Description: Minimum order quantity: 1 unit of 1kit

Cytokeratin 5/8(C-50) Antibody

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EUR 395.00
Description: Primary antibody against Cytokeratin 5/8(C-50), 1mg/mL

Super piggyBac Transposase expression vector (replaces PB200A-1), 50 rxns

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  • Category: PiggyBac Technology

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MR750A-1 50 ul
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  • Category: Stem Cell Products

TR-FRET Bridge-It L-Tryptophan 50

TRF-1-1-1001A 1 Kit
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MTBK 20620 (PE protein)-50

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Aflatoxin G2

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M-phase inducer phosphatase 1 (CDC25A)(MBP-his tagged)-50

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TruStrip Sample Storage/Transportation Trays (5 trays/cluster; 10 clusters for 50 strips) for 15-50 ul strips

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GPK-50 1 kit
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Rv0350 (Heat Shock Protein 70)-50

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Rv2299c (Heat Shock Protein 90)-50

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  • Category: PiggyBac Technology

protein tyrosine phosphatase, non-receptor type 1 (PTPN1)(His tagged)-50

AR09-P0027-50 50ug
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Antigen Retrieval Solution, Citrate pH 6.0 (50 ml, 100X)

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Antigen Retrieval Solution, EDTA pH 8.0 (50 ml, 100X)

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Single channel mini-pipette (130 mm) 50 ul, autoclavable

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PTPRF(His tagged) / LAR(His tagged)-50

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ExoQuick-CG cGMP-grade Exosome Isolation reagent (50 ml)

EXOCG50A-1 50 ml
EUR 1181.00
  • Category: Exosomes

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5-00619 4 x 1mg Ask for price

Adrenomedullin (1-50), rat

HY-P1534 1mg
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Antigen Retrieval Solution, Tris-EDTA pH 9.0 (50 ml, 100X)

ARS9-50 5 L
EUR 286.00

BasicExoRNA? Basic RNA extraction kit (Pre-isolated exosomes, 50 reactions)

K1222-50
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PLGA poly(lactic-co-glycolic) acid); synthetic; (50% lactic and 50% glycolic acid, ~30-60 Kda)

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Adrenomedullin (Adrenomedullin (ADM/AM, 11-50) (rat)

SP-100047-1 1 mg
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slingshot homolog 3 (SSH3)(MBP-his tagged)-50

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AXYPREP MAG DYECLEAN KIT- 50 ML - 5000 PREPS

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Description: Reservoirs; Reservoirs - Axygen

Human IgG/RF-adsorbent (Adsorb 50 serum samples of 100 ul)

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PTPRC(His tagged) / Human CD45 (LCA)(His tagged)-50

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ACP1(MBP-his tagged) / LMPTP(MBP-his tagged)-50

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dual specificity phosphatase 12 (DUSP12)(MBP-his tagged)-50

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515B0 50 mg
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HIV-1 Rev (34-50)

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Description: Sum Formula: C244H361N71O71S6; CAS# [89899-53-6] net

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Description: Sum Formula: C244H361N71O71S6; CAS# [89899-53-6] net

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abx085344-34kDa500mg 3.4 kDa; 500 mg
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Lanosterol, 50%

GP3911-1G 1 g
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Description: Bioscience Mag Beads; Magnetic Fragment Select

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Description: Reservoirs; Reservoirs - Axygen

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dual specificity phosphatase-like 15 (DUSP15)(MBP-his tagged)-50

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dual specificity phosphatase-like 18 (DUSP18)(MBP-his tagged)-50

AR09-P0019-50 50ug
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hSKRP1 mRNA for SKRP1 (NE1, DUSP19)(MBP-his tagged)-50

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dual specificity phosphatase-like 21 (DUSP21)(MBP-his tagged)-50

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dual specificity phosphatase-like 26 (DUSP26)(MBP-his tagged)-50

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protein tyrosine phosphatase zeta-polypeptide (PTPRZ)(MBP-his tagged)-50

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M-phase inducer phosphatase 3 (CDC25C)(MBP-his tagged)-50

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DiagNano Gold Nanorods, diameter 50 nm, absorption max 700 nm

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SureSeal S Sterile Microtube, 1.5mL, packed in self-standing bags of 50, 10 bags/pk

C2000-50 500/pack
EUR 63.16
Description: Micro Centrifuge Tubes

100 bp DNA Ladder Plus, ready-to-use, 50 ug, 0.5 ug per lane

Z6030001-1 100 lanes
EUR 109.00

50 L RPMI 1640, POWDER WITH L-GLUTAMINE, WITHOUT SODIUM BICARBONATE

50-020-PC 50 L/pk
EUR 92.00
Description: Media Catalog; Classical Media

In this study we applied single cell analysis techniques such as flow cytometry (FCM) and fluorescence-assisted cell sorting (FACS) on the spore-forming succinate producer Pseudoclostridium thermosuccinogenes. By combining FCM and FACS with fluorescent staining, we differentiated and enriched all sporulation-related morphologies of P. thermosuccinogenes.

To evaluate the presence of metabolically active vegetative cells, a blend of the dyes propidium iodide (PI) and carboxy fluorescein diacetate (cFDA) tested best. Side scatter (SSC-H) in combination with metabolic indicator cFDA dye provided the best separation of sporulation populations.

Based on this protocol, we successfully determined culture heterogeneity of P. thermosuccinogenes by discriminating between mature spores, forespores, dark and bright phase endospores, and vegetative cells populations. Henceforth, this methodology can be applied to further study sporulation dynamics and its impact on fermentation performance and product formation by P. thermosuccinogenes.

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