N-(Propargyl-PEG2)-PEG3-t-butyl ester is a branched PEG linker with an alkyne group and a t-butyl ester. The alkyne group reacts with azide-bearing compound in coppper catalyzed azide-alkyne Click Chemistry reaction. The amino group is reactive with carboxylic acids, activated NHS esters, and carbonyls. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
N-(Propargyl-PEG2)-PEG3-t-butyl ester is a branched PEG linker with an alkyne group and a t-butyl ester. The alkyne group reacts with azide-bearing compound in coppper catalyzed azide-alkyne Click Chemistry reaction. The amino group is reactive with carboxylic acids, activated NHS esters, and carbonyls. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Isotherm3G is a mesophilic DNA polymerase, perfectly suited for Isothermal amplifications. Isotherm3G has been mutated to have improved reverse transcription activity. It synthesizes DNA from both DNA and RNA templates with a high strand displacement activity allowing simplified one-enzyme RT-LAMP reactions.
Isotherm3G exhibits 5’ to 3’ polymerase activity but lacks any exonuclease activity. The product components have been optimized and are perfectly suited for loop-mediated isothermal amplification (LAMP) with optimal reaction temperature at 65°C.
Isotherm3G DNA Polymerase utilizes an aptamer-based warm-start feature to prevent false amplification at low temperatures. For more information, please contact us.
The product is available also in bulk quantities with custom fillings and can be produced customized for example in a lyo-ready formulation.
Isotherm3G is a mesophilic DNA polymerase, perfectly suited for Isothermal amplifications. Isotherm3G has been mutated to have improved reverse transcription activity. It synthesizes DNA from both DNA and RNA templates with a high strand displacement activity allowing simplified one-enzyme RT-LAMP reactions.
Intended Use
For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials
Principle and Interpretation
Enzymatic digest of casein and enzymatic digest of animal tissue provides nitrogen source, vitamins and growth factors; sodium chloride maintains balanced osmoticpressure; sorbitol is a fermentable sugar; No. 3 bile salt and crystal violet inhibit Gram-positive bacteria, potassiumtellurite inhibits non-o157 Escherichia coli, and cefixime inhibits Proteus; neutral red is a pH indicator; and agar is the coagulant of the culture medium.
Formulation
| Ingredients | /liter |
| Enzymatic digest of casein | 17.0g |
| Enzymatic digest of animal tissue | 3.0g |
| Sorbitol | 10.0g |
| Bile salt No.3 | 1.5g |
| Sodium chloride | 5.0g |
| Neutral red | 0.03g |
| Crystal violet | 0.001g |
| Agar | 15.0g |
| pH 7.1±0.2 at 25°C | |
Preparation
Weigh 51.5g of this product, add 1L of distilled water or deionized water, stir, heat and boil until completely dissolved, dispense into Erlenmeyer flasks, and sterilize at 121℃ for 15min. Melt the sterilized culture medium and cool it to about 45℃, add 1 tube of supporting reagent (SR0240) (0.25mg potassium tellurite and 0.005mg Cefixime) to every 100mL of basal culture medium, mix and pour into the plate.
Quality Control
Cultural characteristics observed after incubation at 35-37°C for 18-24hours
| Quality control strains | Growth | Colony color |
| Escherichia coli ATCC25922 | PR≥0.7 | pale pink to red |
| Escherichia coli 0157:H7 NCTC12900 | PR≥0.7 | colorless |
| Staphylococcus aureus ATCC 25923 | Total inhibition | – |
Sorage and Shelf Life
Keep container tightly closed, store in a cool, dry place, away from bright light. Storage period of 3 years.
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.
Intended Use For the isolation and differentiation of sorbitol-negative Escherichia coli serotype O157 from food and animal feed and other materials Principle and Interpretation Enzymatic ……
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)
Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.
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).
