CO00015 Midstream Cassette 136x16mm

• 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) or allele-specific amplification (ASA).Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.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® Taq 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.HiDi® Taq DNA polymerase harbours a nuclease function and therefor is also suitable for use with hydrolysis probes (TaqMan® probes etc.). It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control and mutation identification in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® Taq 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) or allele-specific amplification (ASA).

Please note: This DNA polymerase is also available as a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.

Description 

YesBlot™ Western Marker I is a ready-to-use mixture with ten IgG-binding proteins covering a wide range of molecular weights from 15 to 200 kDa in Tris-Glycine buffer. YesBlot™ Western Marker I performs dual functions. First, it contains 4 pre-stained proteins (10, 25, 45 and 70 kDa) for monitoring protein separation during SDS-PAGE, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the protein size. Second, ten IgG-binding proteins can be immuno-detected on film or by CCD imaging. YesBlot™ Western Marker I is compatible for chemiluminescent, fluorescent, chromogenic or other detection systems. In addition, YesBlot™ Western Marker I has two reference bands with enhanced intensity (at 30 kDa and 80 kDa). The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading. 

Features

• Direct visualization — 10 IgG-binding proteins for direct visualization on Western blots
• Pre-stained bands — 4 prestained proteins for monitoring protein separation during electrophoresis and Western blotting transferring efficiency on membrane
• Wide range — 10 clear bands from 15 to 200 kDa for size estimation
• Quick reference — two enhanced bands (30 and 80 kDa)

Contents 

The YesBlot™ Western Marker I contains recombinant IgG binding proteins, prestained recombinant proteins, glycerol and SDS. 

Storage

4°C for 3 months
-20°C for 24 months

YesBlot™ Western Marker I is a ready-to-use mixture with ten IgG-binding proteins covering a wide range of molecular weights from 15 to 200 kDa in Tris-Glycine buffer. YesBlot™ Western Marker I performs dual functions. First, it contains 4 pre-stained proteins (10, 25, 45 and 70 kDa) for monitoring protein separation during SDS-PAGE, verification of Western transfer efficiency on membranes (nitrocellulose, PVDF, or nylon) and for approximating the protein size. Second, ten IgG-binding proteins can be immuno-detected on film or by CCD imaging. YesBlot™ Western Marker I is compatible for chemiluminescent, fluorescent, chromogenic or other detection systems. In addition, YesBlot™ Western Marker I has two reference bands with enhanced intensity (at 30 kDa and 80 kDa). The marker is supplied in the gel loading buffer and is ready to use. Do NOT heat, dilute, or add reducing agents before loading.

Product Details

Application:

DNA Nucleic Acid

Format:

NFO Kit

Kit Components:

Reagents,Buffer,Enzymes

Manufacturer:

Amp-future Bio

Product Name:

DNA Isothermal Rapid Amplification Kit (Colloidal Gold Test Strip Type)

Reaction Time:

5-20mins

Reaction Volume:

50μL

Sensitivity:

500-1000copies/ml

Shelf Life:

14 Months

Specificity:

High

Storage Conditions:

-20℃

Suitability:

Universal

High Light:

Isothermal DNA Amplification Kit

, 

DNA Amplification Kit NFO

, 

20mins nucleic acid amplification

Payment & Shipping Terms

Minimum Order Quantity

48T

Price

USD$3.8/T

Packaging Details

16T/Bag,48T/Box

Delivery Time

6 working days

Payment Terms

T/T, MoneyGram

Supply Ability

100000T/Month

Product Description

DNA Isothermal Rapid Amplification Kit (Colloidal Gold Test Strip Type)

【Principle Overview】

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins, the recombinase and primers form a complex; Source search and combine the target homology domain, at this time, a D-loop region is formed at the homology position and strand exchange begins; along with the dissociation of the recombinase from the complex, the polymerase also binds to the 3′ end of the primer and begins chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.

【Primer design】

It is recommended to use primers with a length of 30-35 bp.Too short primers will affect the amplification speed and detection sensitivity;the 5′ end of the reverse primer is labeled with a modification group (commonly used biotin).

Primers are designed to avoid the formation of secondary structures that affect amplification; the length of the amplicon is recommended to be 150-500bp.

【Colloidal gold probe design】

In the middle of the forward and reverse primer,design a sequence of 46-52nt in length that is complementary to the target fragment; modify an antigen marker (typical FAM) at the 5′ end;mark a dSpacer (tetrahydrofuran, THF) in the middle of the 5′ end and the 3′ end ), as the recognition site of nfo;the 3′ end is labeled with a modification group, such as amine group, phosphate group or C3-Spacer, etc.

【Features】

This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 15 minutes),and the reaction components are in dry powder state,which is easy to operate and easy to store.

This reagent has low equipment requirement,and the reaction operation can be carried out in metal baths, water baths, etc.,and there is no need to purchase expensive exclusive equipment such as PCR amplifiers.

【Kit storage】

1. Storage conditions: storage temperature ≤ -20℃constant temperature environment, keep away from light and avoid heavy pressure;

2. Product validity period: 14 months;

3. See the outer packaging for the production date.

This kit is based on a rapid nucleic acid amplification technology at room temperature and constant temperature: at room temperature and constant temperature (generally 39℃~42℃), with the help of auxiliary proteins and single-strand binding proteins, the recombinase and primers form a complex; Source search and combine the target homology domain, at this time, a D-loop region is formed at the homology position and strand exchange begins; along with the dissociation of the recombinase from the complex, the polymerase also binds to the 3′ end of the primer and begins chain extension. Relying on the action of nfo enzyme, adding specific molecular probes designed according to the template, and using colloidal gold technology (sandwich method) can detect the final result.