Kbc stain

Manufactured by CinnaGen

The KBC stain is a laboratory reagent used for the staining and visualization of biological samples. It serves as a general-purpose stain, allowing for the identification and differentiation of various cellular structures and components. The KBC stain's core function is to provide clear and consistent staining, enabling researchers and technicians to analyze and interpret their samples more effectively.

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Lab products found in correlation

Thermal cycler, by Analytik Jena (1 mentions) Dna polymerase master mix red, by Ampliqon (1 mentions) Veriti 96 well thermal cycler, by Thermo Fisher Scientific (1 mentions) Taq polymerase, by CinnaGen (1 mentions)

3 protocols using kbc stain

Detecting Carbapenemase Genes in Pseudomonas aeruginosa

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The imipenem-resistant isolates of P. aeruginosa were selected. Next, their total DNA was extracted by the boiling method [13 (link),14 (link)] and stored at -20°C until PCR amplification. Polymerase chain reaction (PCR) was carried out to detect blaIMP and blaVIM genes and common subtypes of bla_IMP-1, bla_IMP-2, bla_VIM-1, and bla_VIM-2 with specific primers (Table 1). The gels were stained with KBC stain (CinnaGen, Iran), and the PCR products were visualized with UV light [10 (link)].Table 1

primers sequence of target genes

Table 1

Target gene	Primer	Oligonucleotide sequence (5›–3›)	size (pb)
VIM	blaVIM	Forward: GAT GGT GTT TGG TCG CAT A	390
VIM	blaVIM	Reverse: CGA ATG CGC AGC ACC AG	390
IMP	blaIMP	Forward: GGA ATA GAG TGG CTT AAY TCT C	232
IMP	blaIMP	Reverse: GGT TTA AYA AAA CAA CCA CC	232
VIM1	blaVIM1	Forward: AGT GGT GAG TAT CCG ACA G	261
VIM1	blaVIM1	Reverse: ATG AAA GTG CGT GGA GAC	261
VIM2	blaVIM2	Forward: ATG TTC AAA CTT TTG AGT AAG	801
VIM2	blaVIM2	Reverse: CTA CTC AAC GAC TGA GCG	801
IMP1	blaIMP1	Forward: ACC GCA GCA GAG TCT TTG CC	586
IMP1	blaIMP1	Reverse: ACA ACC AGT TTT GCC TTA CC	586
IMP2	blaIMP2	Forward: GTT TTA TGT GTA TGC TTC C	673
IMP2	blaIMP2	Reverse: AGC CTG TTC CCA TGT AC	673

Karampoor M., Akhlaghi F., Mobayen M.R., Afrasiabi F., Khodayary R., Moradzadeh M, & Nikokar I. (2022). Phenotypic and genotypic characterization of metallo-β-lactamase producing Pseudomonas aeruginosa isolated from burn patients. New Microbes and New Infections, 49-50, 101059.

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Molecular Detection of Carbapenemase Genes

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The boiling method was applied to extract the template DNA to detect carbapenemase genes. The primers were used to detect bla_KPC, bla_OXA-48-like, bla_IMP, bla_VIM, and bla_NDM as described by Doyle et al.5 (link)
The amplification reaction was carried out in a final volume of 25 μL containing 0.5 μL of each primer (10 pM), 12.5 μL of DNA Polymerase Master Mix RED (Ampliqon, Co., Denmark), 1 μL of DNA, and 10.5 μL of DNase and RNase-free water in a FlexCycler PCR. The Thermal Cycler (Analytik Jena, Germany) was performed through the following cycling conditions: an initial denaturation step at 95 °C for 5 min, followed by 35 cycles of DNA denaturation at 95 °C for 45 s, annealing at 45 s, and extension at 72 °C for 1 min, and a final extension at 72 °C for 8 min.
The PCR products were analyzed with gel electrophoresis on a 1.5% agarose gel in 0.5 Tris, EDTA, and Boric acid (TBE) buffer, followed by staining with KBC stain (CinnaGen, Iran) and visualized using a UV transilluminator. Positive controls for these genes were obtained from Pasteur Institute of Iran, Tehran.

Hashemizadeh Z., Hosseinzadeh Z., Azimzadeh N, & Motamedifar M. (2020). Dissemination Pattern of Multidrug Resistant Carbapenemase Producing Klebsiella pneumoniae Isolates Using Pulsed-Field Gel Electrophoresis in Southwestern Iran. Infection and Drug Resistance, 13, 921-929.

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Molecular Detection of Carbapenemase Genes

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DNA extraction from the studied isolates was done by boiling method, as described previously. 4 PCR amplification for detection of bla KPC , bla OXA-48-like, bla NDM , bla VIM , and bla IMP genes was carried out on a Veriti 96-well thermal cycler instrument (Applied Biosystems at Life Technologies, Foster City, CA), as described previously. 10 The primers were used at concentrations of 0.3-0.4 μM. They were provided by Takapoo zist Co, Tehran, Iran. The primer sequences and amplicon sizes are listed in Table 1. Amplification reaction was performed in a final volume of 25 μL containing 200 μM concentrations of dNTPs, 1.5 mM MgCl 2 , 2 U taq polymerase (CinnaGen, Iran) and 3 μL DNA templates. Positive controls for targeted genes were kindly obtained from Pasteur Institute, Tehran, Iran. The PCR program consisted of an initial denaturation step at 95 °C for 5 min, followed by 35 cycles of DNA denaturation at 95 °C for 45 s, primer annealing for 45 s(Temperature was depending on the sequences of primers), and primer extension at 72 °C for 1 min, followed by a final extension at 72 °C for 8 min. The PCR products were analyzed by electrophoresis on 1.5% agarose gels in 0.5 × Tris-Acetate-EDTA (TAE) buffer. The gels were stained with KBC stain (CinnaGen, Iran) and the PCR products were visualized with UV light.

Hosseinzadeh Z., Sedigh Ebrahim-Saraie H., Sarvari J., Mardaneh J., Dehghani B., Rokni-Hosseini S.M.H, & Motamedifar M. (2018). Emerge of bla_NDM-1 and bla_OXA-48-like harboring carbapenem-resistant Klebsiella pneumoniae isolates from hospitalized patients in southwestern Iran. Journal of the Chinese Medical Association : JCMA, 81(6).

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