Joseph Grant (University of East Anglia)

Biomathematics, Biostatistics,Genetics, Molecular Biology

Molecular Biology Methods in Streptomyces rimosus

Molecular Biology Methods in Streptomyces rimosus

Molecular Biology Strategies in Streptomyces rimosus, a Producer of Oxytetracycline

Streptomyces rimosus is used for manufacturing of the broad-spectrum antibiotic oxytetracycline (OTC). S. rimosus belongs to Actinomyces species, a big group of microorganisms that produce various set of pure metabolites of excessive significance in lots of points of our life. On this chapter, we describe particular molecular biology strategies and a classical homologous recombination method for focused in-frame deletion of a goal gene or complete operon in S. rimosus genome. The introduced protocols will information you thru the design of experiment and building of homology arms and their cloning into acceptable vectors, that are appropriate for gene-engineering work with S. rimosus. Moreover, two totally different protocols for S. rimosus transformation are described together with detailed process for focused gene substitute by way of double crossover recombination occasion.

Gene deletion is confirmed by colony PCR, and colonies are additional characterised by cultivation and metabolite evaluation. As the ultimate step, we current in trans complementation of the deleted gene, to verify performance of the engineering method achieved by gene disruption. A lot of methodological steps and protocols are optimized for S. rimosus strains together with the usage of the chosen reporter genes. Protocols described on this chapter may be utilized for learning operate of any particular person gene product in various OTC-producing Streptomyces rimosus strains.

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Casein, for Molecular Biology
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Sodium Chloride for molecular biology, 99.9%
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Isopropanol (IPA) for molecular biology, 99.8%
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Acetonitrile (ACN) for molecular biology, 99.9%
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Phenol Crystalline for molecular biology, 99.5%
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Sodium Bicarbonate for molecular biology, 99.7%
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Glycerol (Glycerine) for molecular biology, 99.5%
62417 100 ml
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Potassium Chloride for molecular biology, 99.5%
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Dichloromethane (DCM) for molecular biology, 99.9%
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Acriflavine Neutral for molecular biology, 98%
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2-Mercaptoethanol for molecular biology, 99%
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Tetrahydrofuran (THF) for molecular biology, 99.8%
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Dithioerythritol (DTE) for molecular biology, 99%
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meta-Topolin (mT) for molecular biology, 98%
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Glycogen ex. Oyster for molecular biology, 85%
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Propidium Iodide (PI) for molecular biology, 95%
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Acrylamide 3x cryst. for molecular biology, 99.9%
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D-Sorbitol Powder for molecular biology, 98%
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Acriflavine Hydrochloride for molecular biology, 98.5%-105%
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Tris Buffer Superior for molecular biology, 99.9%
37969 100 Gms
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Polysorbate 20 (Tween 20) for molecular biology
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Molecular Biology Grade Water for RT-PCR
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Description: Molecular Biology Grade Water for RT-PCR

10X TE Buffer pH-8.0 for molecular biology
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Tris Buffer AR, ACS for molecular biology, 99.9%
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10X TBE Buffer pH-8.3 for molecular biology
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Acrylamide 40% aq. solution for molecular biology
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DL-Dithiothreitol (DTT) for molecular biology, 98%
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Glutathione Oxidized (GSSG) for molecular biology, 99.5%
22151 250 Mg
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Cesium Chloride ultrapure for molecular biology, 99.9%
32355 25 Gms
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Guanidine Thiocyanate (GTC) for molecular biology, 99%
80272 100 Gms
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88035 250 Gms
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Pyridine, GlenBiol™, suitable for molecular biology with molecular sieve
GS8780 2500ml
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46791 5 ml
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Pyridine, GlenBiol™, suitable for molecular biology
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Formamide, GlenBiol™, suitable for molecular biology
GS9663 100ml
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Bromocresol Purple Sodium Salt for molecular biology
24836 5 Gms
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Guanidine Hydrochloride (GHC) for molecular biology, 99.5%
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Description: Part A

20X SSC Buffer pH 6.9-7.1 for molecular biology
12590 200 ml
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50488 100 Gms
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24017 100 ml
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57806 200 ml
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OORA00229-1L - Molecular Biology Grade UltraPure Water
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OORA00230-1L - Molecular Biology Grade UltraPure Water
OORA00230-1L 1L
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DTT (Molecular Biology Grade)
CE131 5 g
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DTT (Molecular Biology Grade)
CE132 10 g
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CE133 25 g
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CE196 1 g
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CE197 5 g
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NBT (Molecular Biology Grade)
CE209 1 g
EUR 123.6

Past descriptive accuracy: The central dogma of molecular biology in scientific follow

There isn’t a denying the Central Dogma’s influence on the organic sciences. Because the Dogma’s formulation by Francis Crick in 1958, nonetheless, many have debated the Dogma’s empirical adequacy. My goal is to maneuver past these discussions, and as a substitute take into account the Central Dogma’s significance to modern organic follow.

To do that, I take into account 4 distinct approaches for figuring out the non-descriptive methodological significance of a scientific precept. I argue that these approaches fail to vindicate the Central Dogma, and that, below many of those approaches, the Dogma quantities to a triviality.

Molecular Biology of Bladder Most cancers: Potential Implications for Remedy

Lately accomplished research offered high-resolution descriptions of the molecular organic traits of urothelial bladder cancers. Complete transcriptome messenger RNA expression profiling revealed that they are often grouped into basal and luminal molecular subtypes resembling those described in breast cancers.

Retrospective DNA sequencing efforts revealed roles for disruption of DNA injury response pathways in response to standard chemotherapy and immune checkpoint blockade, and accomplished and ongoing research point out that the molecular organic properties of infiltrating host cells dictate additionally affect therapeutic outcomes. This text opinions these findings and establish gaps in information that characterize alternatives for future analysis.

A guided-inquiry investigation of genetic variants utilizing Oxford nanopore sequencing for an undergraduate molecular biology laboratory course

Subsequent Era Sequencing (NGS) has grow to be an necessary instrument within the organic sciences and has a rising variety of purposes throughout medical fields. Presently, few undergraduate packages present coaching within the design and implementation of NGS purposes. Right here, we describe an inquiry-based laboratory train for a college-level molecular biology laboratory course that makes use of real-time MinION deep sequencing and bioinformatics to research attribute genetic variants present in most cancers cell-lines. The general objective for college students was to establish non-small cell lung most cancers (NSCLC) cell-lines primarily based on their distinctive genomic profiles.

The models described on this laboratory spotlight core rules in multiplex PCR primer design, real-time deep sequencing, and bioinformatics evaluation for genetic variants. We discovered that the MinION machine is an acceptable, possible instrument that gives a complete, hands-on NGS expertise for undergraduates. Pupil evaluations demonstrated elevated confidence in utilizing molecular methods and enhanced understanding of NGS ideas. General, this train gives a pedagogical instrument for incorporating NGS approaches within the educating laboratory as manner of enhancing college students’ comprehension of genomic sequence evaluation. Additional, this NGS lab module can simply be added to a wide range of lab-based programs to assist undergraduate college students be taught present DNA sequencing strategies with restricted effort and price.

Prioritizing Molecular Biomarkers in Bronchial asthma and Respiratory Allergy Utilizing Programs Biology

Extremely prevalent respiratory ailments reminiscent of bronchial asthma and allergy stay a urgent well being problem. Presently, there may be an unmet want for exact diagnostic instruments able to predicting the nice heterogeneity of those diseases. In a earlier research of 94 bronchial asthma/respiratory allergy biomarker candidates, we outlined a bunch of potential biomarkers to differentiate medical phenotypes (i.e. nonallergic bronchial asthma, allergic bronchial asthma, respiratory allergy with out bronchial asthma) and illness severity. Right here, we analyze our experimental outcomes utilizing advanced algorithmic approaches that set up holistic illness fashions (programs biology), combining these insights with info obtainable in specialised databases developed worldwide. With this method, we goal to prioritize probably the most related biomarkers in keeping with their specificity and mechanistic implication with molecular motifs of the ailments. The Therapeutic Efficiency Mapping System (Anaxomics’ TPMS know-how) was used to generate one mathematical mannequin per illness: allergic bronchial asthma (AA), non-allergic bronchial asthma (NA), and respiratory allergy (RA), defining particular molecular motifs for every.

The connection of our molecular biomarker candidates and every illness was analyzed by synthetic neural networks (ANNs) scores. These analyses prioritized molecular biomarkers particular to the ailments and to explicit molecular motifs. As a primary step, molecular characterization of the pathophysiological processes of AA outlined 16 molecular motifs: 2 particular for AA, 2 shared with RA, and 12 shared with NA. Mechanistic evaluation confirmed 17 proteins that had been strongly associated to AA. Eleven proteins had been related to RA and 16 proteins with NA. Specificity evaluation confirmed that 12 proteins had been particular to AA, 7 had been particular to RA, and a couple of to NA. Lastly, a triggering evaluation revealed a related function for AKT1, STAT1, and MAPK13 in all three situations and for TLR4 in asthmatic ailments (AA and NA). In conclusion, this research has enabled us to prioritize biomarkers relying on the performance related to every illness and with particular molecular motifs, which may enhance the definition and usefulness of recent molecular biomarkers.

Molecular Biology of Ovarian Most cancers: From Mechanisms of Intraperitoneal Metastasis to Therapeutic Alternatives

Ovarian most cancers (OC) is likely one of the most frequent malignancies of the feminine genital tract, and continues to be the main explanation for loss of life from gynecological tumors […].

Guanidine Thiocyanate (Molecular Biology Grade)
CE164 100 g
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Guanidine Thiocyanate (Molecular Biology Grade)
CE165 500 g
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Guanidine Thiocyanate (Molecular Biology Grade)
CE166 1 kg
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Tris - Hydrochloride (Molecular Biology Grade)
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Guanidine Hydrochloride
09860-25 500G
EUR 224

Guanidine Hydrochloride
17318-24 10KG
EUR 742

Guanidine Hydrochloride
17318-82 25G
EUR 14

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17318-95 500G
EUR 68.6

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EUR 32.2

Guanidine Hydrochloride
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17353-25 500G
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18-206 500g/Unit
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40700060-2 500 g
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40700060-3 1 kg
EUR 153.37

Guanidine Hydrochloride
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A2719-500G 500G
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Guanidine Hydrochloride
G821245 10g
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Guanidine hydrochloride
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Guanidine thiocyanate?For Molecular Biol
MB015-1KG 1 unit
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Description: Guanidine thiocyanate?For Molecular Biol

Guanidine thiocyanate?For Molecular Biol
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Guanidine hydrochloride, 99%
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G821249 25mg
EUR 1200
Description: 121616-39-5

Guanidine-d5 (hydrochloride)
HY-B0178AS1 Get quote Ask for price
Description: Guanidine-d5 (hydrochloride) is the deuterium labeled Guanidine hydrochloride[1]. Guanidine hydrochloride (Guanidinium chloride) a strong chaotrope, is also a strong denaturant of proteins[2][3].

Guanidine hydrochloride solution, 8M
GE1001 100ml
EUR 386.21

Guanidine hydrochloride solution, 6M
GE4739 100ml
EUR 651.82

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EUR 485

Guanidine-13C,15N3 (hydrochloride)
HY-B0178AS 1mg Ask for price
Description: Guanidine-13C,15N3 (hydrochloride)is the 13C-labeled and 15N-labeled Guanidine hydrochloride. Guanidine hydrochloride (Guanidinium chloride) a strong chaotrope, is also a strong denaturant of proteins[1][2].

Guanidine hydrochloride, Hi-LR™
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Description: Guanidine hydrochloride, Hi-LR™

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Description: Guanidine hydrochloride, Hi-LR™

Guanidine Hydrochloride (GHC) pure, 99%
54542 100 Gms
EUR 3.01
Description: Part A

Guanidine hydrochloride (6 M) solution
B1013-1L each
EUR 301.2

Guanidine hydrochloride (6 M) solution
B1013-4L each
EUR 705.6

1-(2,4-difluorophenyl)guanidine hydrochloride
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Description: 1-(2,4-difluorophenyl)guanidine hydrochloride

1-(2,4-difluorophenyl)guanidine hydrochloride
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Description: 1-(2,4-difluorophenyl)guanidine hydrochloride

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Description: 1-(2,4-difluorophenyl)guanidine hydrochloride

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Description: 1-(2,4-difluorophenyl)guanidine hydrochloride

1-(2,4-difluorophenyl)guanidine hydrochloride
T50030-5mg 5mg Ask for price
Description: 1-(2,4-difluorophenyl)guanidine hydrochloride

1-(2,4-difluorophenyl)guanidine hydrochloride
MBS5785869-10mg 10mg
EUR 180

1-(2,4-difluorophenyl)guanidine hydrochloride
MBS5785869-25mg 25mg
EUR 235

1-(2,4-difluorophenyl)guanidine hydrochloride
MBS5785869-50mg 50(mg
EUR 300

1-(2,4-difluorophenyl)guanidine hydrochloride
MBS5785869-5mg 5(mg
EUR 155

N-(3,4-Dimethoxyphenyl)guanidine Hydrochloride
D470625 10g
EUR 800
Description: 855890-38-9

N-Cyano-N'-(6-guanidinohexyl) Guanidine Hydrochloride
C655610 250mg
EUR 400

N-​(4-​Methyl-​2-​thiazolyl)​-guanidine hydrochloride
M331435 100mg
EUR 58
Description: 100599-91-5

N-(6-Aminohexyl)-N'-(6-guanidinohexyl) Guanidine Hydrochloride
A544590 50mg
EUR 6000

Casein, for Molecular Biology
MB279-100G 1 unit
EUR 110.77
Description: Casein, for Molecular Biology

Casein, for Molecular Biology
MB279-500G 1 unit
EUR 387.65
Description: Casein, for Molecular Biology

Sucrose for molecular biology
27580 500 Gms
EUR 4.11
Description: Part A

1-(4-Chloromethyl-2-thiazoyl)guanidine Hydrochloride Salt
C369545 1g
EUR 146

1-(4-Chloromethyl-2-thiazoyl)guanidine Hydrochloride Salt
MBS6102657-5g 5(g
EUR 515

1-(4-Chloromethyl-2-thiazoyl)guanidine Hydrochloride Salt
MBS6102657-5x5g 5x5g
EUR 2175

Urea for molecular biology, 99.5%
21113 500 Gms
EUR 6.5
Description: Part A

Xylene for molecular biology, 99.5%
45122 250 ml
EUR 2.81
Description: Part A

Acetone for molecular biology, 99.8%
27498 500 ml
EUR 3.76
Description: Part A

Glycine for molecular biology, 99.5%
64072 100 Gms
EUR 1.64
Description: Part A

Carbinol for molecular biology, 99.5%
34883 500 ml
EUR 6.23
Description: Part A

Methanol for molecular biology, 99.5%
96446 500 ml
EUR 6.23
Description: Part A

Formamide for molecular biology, 99.5%
30349 500 ml
EUR 5.82
Description: Part A

1-(4-Chloromethyl-2-thiazoyl)guanidine-13C3 Hydrochloride Salt
MBS6089660-1mg 1(mg
EUR 605

1-(4-Chloromethyl-2-thiazoyl)guanidine-13C3 Hydrochloride Salt
MBS6089660-5x1mg 5x1mg
EUR 2565

Imidazole for molecular biology, 99.5%
61510 25 Gms
EUR 5.47
Description: Part A

Chloroform for molecular biology, 99.8%
96764 100 ml
EUR 3.08
Description: Part A

Urea, suitable for molecular biology
GE1210 500g
EUR 53.1

Urea, suitable for molecular biology
GE1210-1 1
EUR 58

Urea, suitable for molecular biology
GE1210-1KG 1 kg
EUR 106.8

Urea, suitable for molecular biology
GE1210-500 500
EUR 33.1

Urea, suitable for molecular biology
GE1210-500G 500 g
EUR 76.8

Triethylamine for molecular biology, 99.5%
55221 500 ml
EUR 4.11
Description: Part A

Ficoll 400® for molecular biology
45460 5 Gms
EUR 15.6
Description: Part B

Agarose UltraPhor for molecular biology
27813 10 Gms
EUR 46.5
Description: Part B

Boric Acid for molecular biology, 99.5%
22311 250 Gms
EUR 4.04
Description: Part A

2-Mercaptoethanol ?For Molecular Biology
MB041-100ML 1 unit
EUR 9.02
Description: 2-Mercaptoethanol ?For Molecular Biology

2-Mercaptoethanol ?For Molecular Biology
MB041-500ML 1 unit
EUR 26.47
Description: 2-Mercaptoethanol ?For Molecular Biology

ACES Buffer for molecular biology, 99.5%
27907 5 Gms
EUR 5.4
Description: Part B

Triton X-100 for molecular biology
64518 100 ml
EUR 6.02
Description: Part A

MOPS Buffer for molecular biology, 99%
66043 25 Gms
EUR 10.28
Description: Part B

Vitamin B12 for molecular biology, 97%
77472 250 Mg
EUR 5.78
Description: Part B

N-[2-[(6-Amino-5-nitro-2-pyridinyl)amino]ethyl]guanidine Hydrochloride
MBS6111956-10mg 10(mg
EUR 545

N-[2-[(6-Amino-5-nitro-2-pyridinyl)amino]ethyl]guanidine Hydrochloride
MBS6111956-5x10mg 5x10mg
EUR 2310

HEPES Buffer for molecular biology, 99.5%
16826 25 Gms
EUR 6.3
Description: Part B

Sodium Azide for molecular biology, 99%
17782 100 Gms
EUR 5.47
Description: Part A

CHAPS Buffer for molecular biology, 99.5%
21420 1 Gms
EUR 10.73
Description: Part B

Agarose Low EEO for molecular biology
36601 10 Gms
EUR 5.1
Description: Part B

PIPES Buffer for molecular biology, 99%
49159 100 Gms
EUR 26.03
Description: Part B

10X MOPS Buffer for molecular biology
68078 100 ml
EUR 4.5
Description: Part B

Agarose High EEO for molecular biology
60645 25 Gms
EUR 13.8
Description: Part B

Jasmonic Acid for molecular biology, 95%
79238 25 Mg
EUR 120.9
Description: Part B

Protamine Sulfate for molecular biology, 90-110%
78349 1 Gms
EUR 22.5
Description: Part B

Oil Red O for molecular biology, 75%
23576 25 Gms
EUR 5.33
Description: Part B

Agarose Medium EEO for molecular biology
10423 10 Gms
EUR 6.75
Description: Part B

Sodium Chloride for molecular biology, 99.9%
33205 500 Gms
EUR 4.52
Description: Part A

Isopropanol (IPA) for molecular biology, 99.8%
38445 100 ml
EUR 1.71
Description: Part A

Agarose Low Melting for molecular biology
32417 5 Gms
EUR 22.5
Description: Part B

Ammonium Acetate for molecular biology, 98%
37829 100 Gms
EUR 2.12
Description: Part A

Acetonitrile (ACN) for molecular biology, 99.9%
62006 250 ml
EUR 4.93
Description: Part A

Ethidium Bromide for molecular biology, 95%
93079 1 Gms
EUR 4.58
Description: Part A

Ammonium Chloride for molecular biology, 99.5%
16992 500 Gms
EUR 4.52
Description: Part A

Ammonium Sulphate for molecular biology, 99.5%
82126 250 Gms
EUR 4.93
Description: Part A

Potassium Acetate for molecular biology, 99.5%
96248 100 Gms
EUR 2.74
Description: Part A

Phenol Crystalline for molecular biology, 99.5%
17286 100 Gms
EUR 3.15
Description: Part A

Sodium Bicarbonate for molecular biology, 99.7%
36328 500 Gms
EUR 5.47
Description: Part A

Glycerol (Glycerine) for molecular biology, 99.5%
62417 100 ml
EUR 2.05
Description: Part A

Potassium Chloride for molecular biology, 99.5%
84984 500 Gms
EUR 6.16
Description: Part A

Dichloromethane (DCM) for molecular biology, 99.9%
41512 250 ml
EUR 3.22
Description: Part A

Acriflavine Neutral for molecular biology, 98%
42428 5 Gms
EUR 3.15
Description: Part B

2-Mercaptoethanol for molecular biology, 99%
83759 100 ml
EUR 5.27
Description: Part A

Tetrahydrofuran (THF) for molecular biology, 99.8%
92562 250 ml
EUR 3.28
Description: Part A

Dithioerythritol (DTE) for molecular biology, 99%
53384 1 Gms
EUR 9.53
Description: Part B

Sodium chloride, suitable for molecular biology
GE0307 1kg
EUR 41.33

Sodium chloride, suitable for molecular biology
GE0307-1 1
EUR 45.2

L-Ascorbic Acid for molecular biology, 99.7%
14116 25 Gms
EUR 1.64
Description: Part A

HEPES Sodium Salt for molecular biology, 99%
32225 25 Gms
EUR 11.85
Description: Part B

meta-Topolin (mT) for molecular biology, 98%
68913 25 Mg
EUR 17.48
Description: Part B

Sucrose, GlenBiol, suitable for molecular biology
GC3201-1KG 1 kg
EUR 90

Glycogen ex. Oyster for molecular biology, 85%
49740 1 Gms
EUR 7.5
Description: Part B

Propidium Iodide (PI) for molecular biology, 95%
11195 10 Mg
EUR 17.63
Description: Part B

Acetic Acid Glacial for molecular biology, 99.9%
59788 100 ml
EUR 3.56
Description: Part A

Acrylamide 3x cryst. for molecular biology, 99.9%
61346 100 Gms
EUR 5.47
Description: Part A

Brilliant Blue G-250 for molecular biology
64222 5 Gms
EUR 4.65
Description: Part B

Brilliant Blue R-250 for molecular biology
93473 5 Gms
EUR 4.35
Description: Part B

D-Sorbitol Powder for molecular biology, 98%
99103 500 Gms
EUR 10.33
Description: Part A

Tris Buffer Superior for molecular biology, 99.9%
37969 100 Gms
EUR 7.59
Description: Part A

Polysorbate 20 (Tween 20) for molecular biology
65296 100 Gms
EUR 10.81
Description: Part A

Molecular Biology Grade Water for RT-PCR
ML065-1.5ML 1 unit
EUR 7.82
Description: Molecular Biology Grade Water for RT-PCR

10X TE Buffer pH-8.0 for molecular biology
51782 100 ml
EUR 9.45
Description: Part B

10X TAE Buffer pH-8.3 for molecular biology
71989 100 ml
EUR 4.43
Description: Part B

Tris Buffer AR, ACS for molecular biology, 99.9%
71033 100 Gms
EUR 6.02
Description: Part A

10X TBE Buffer pH-8.3 for molecular biology
83170 200 ml
EUR 4.95
Description: Part B

MES Monohydrate Buffer for molecular biology, 99%
26854 25 Gms
EUR 11.03
Description: Part B

Acrylamide 40% aq. solution for molecular biology
29787 250 ml
EUR 20.53
Description: Part A

Glutathione Reduced (GSH) for molecular biology, 99.5%
11514 1 Gms
EUR 5.55
Description: Part B

DL-Dithiothreitol (DTT) for molecular biology, 98%
17315 1 Gms
EUR 5.18
Description: Part B

Sodium Acetate Anhydrous for molecular biology, 99%
22342 250 Gms
EUR 3.56
Description: Part A

Dimethyl Sulphoxide (DMSO) for molecular biology, 99.8%
24075 100 ml
EUR 6.02
Description: Part A

Trifluoroacetic Acid (TFA) for molecular biology, 99.9%
40801 100 ml
EUR 22.92
Description: Part A

Ammonium Persulphate (APS) for molecular biology, 99%
65553 25 Gms
EUR 1.64
Description: Part A

Glutathione Oxidized (GSSG) for molecular biology, 99.5%
22151 250 Mg
EUR 22.5
Description: Part B

Cesium Chloride ultrapure for molecular biology, 99.9%
32355 25 Gms
EUR 18
Description: Part B

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