GeneAb™ SOX10

Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.

Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

Introduction

Intended Use

For selective enrichment culture of Listeria monocytogenes in food.

Principle and Interpretation

Tryptone, proteose peptone, meat extract, yeast paste powder provide nitrogen sources, vitamins, amino acids, and growth factors; sodium chloride can maintain a balanced osmotic pressure; sodium dihydrogen phosphate and potassium dihydrogen phosphate as buffering agents; esculin are fermentable sugars; lithium chloride and other antibiotics can inhibit Gram negative bacteria and most Gram positive bacteria.

Formulation

Ingredients	/liter
Enzymatic digest of animal tissues　( Proteose peptone)	5.0g
Enzymatic digest of casein　( Tryptone)	5.0g
Meat extract	5.0g
Yeast extract	5.0g
Sodium chloride	20.0g
Disodium hydrogen phosphate dihydrate	12.0g
Potassium dihydrogen phosphate	1.35g
Aesculin	1.0g
Lithium chloride	3.0g
pH7.2±0.2 at 25°C

Preparation

Suspend 57.4g in 1 L of distilled or deionized water. Heat with frequent agitation and boil to completely dissolve the powder. Distribute into flasks. Autoclave at 121℃ for 15 minutes. Cool to 45-50℃,aseptically add the contents of 1 vial of Fraser Selective Supplement (SR0120) to each 225 mL of base to prepare FB1 solution; Add one reagent (SR0130) to each100 mL of FB2 culture medium to prepare FB2 enrichment solution.

Quality Control

Cultural characteristics observed after incubation at 35-37°C for 46~50 hours

Quality control strains	Growth	Characteristics
Listeria monocytogenes ATCC19115 + Escherichia coli ATCC25922 + Enterococcus faecalis ATCC29212	>20 cfu  On PALCAM	Gray to black colony count with black halo
Escherichia coli ATCC25922	< 100 colonies on TSA	Inhibition
Enterococcus faecalis ATCC29212

Storage and Shelf Life

2-30℃，Keep container tightly closed, avoid direct sunlight.

Use before expiry date on the label.

 Precautions

1. When weighing the dehydrated medium, please wear masks to avoid causing respiratory system discomfort

2. Keep container tightly closed after using to prevent clumping.

Waste Disposal

Microbiological contamination was disposed by autoclaving at 121°C for 30 minutes.

Revision

On June 14, 2024

References

ISO 11290

Intended Use For selective enrichment culture of Listeria monocytogenes in food. Principle and Interpretation Tryptone, proteose peptone, meat extract, yeast paste powder provide nitrogen ……

• HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).This polymerase is also available as a full-length Taq DNA polymerase with a nuclease domain, featuring 100% compatibility with hydrolysis probes (TaqMan® probes etc.).Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.Several independently conducted studies show that HiDi® DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)
  For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.

HiDi is available as:

HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)

Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.

Example Results – There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.

HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).