Boc-Gly-PEG3-endo-BCN

• DirectBlood Genotyping PCR Kit is a novelty on the market. It allows genotyping in real-time and without any previous DNA isolation. You simply save time and money: directly use EDTA blood without any extraction steps.
  The DirectBlood Genotyping PCR Kit is optimized to function with a wide range of different SNPs to detect possible nucleotide variations. It has been successfully tested with a variety of hydrolysis probes and hybridization probes.The setup of a genotyping experiment can not be simpler anymore: Mix primers&probes with the Genotying mastermix, add the respective blood sample and start real-time PCR, this is it. 
  
  It is even more simple: store the DirectBlood Genotyping PCR Kit at room-temperature. A cool chain is not needed anymore. The included master mix is freeze-dried and dissolves within a few seconds after addition of the rehydration buffer.Additionally, we have already established SNP-assays for you. In case, you can not find your respective SNPs, please provide us with the rs-number of the SNP-assays your are interested-in and you will receive an individual quotation from us.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.

DirectBlood Genotyping Kit (#5000) performance with a hydrolysis probe assay

The DirectBlood Genotyping Kit is compatible with a wide range of assays: Here we are detecting a SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hydrolysis probe assay.

Homozygous wild type blood samples will show a strong signal and amplification plot (blue curve) in channel 2 (Cy5) and none or a very low signal

(red curve) in the other channel 1 (ROX).

Heterozygous blood samples will give an intermediate signal and amplification plots with very similar intensities in both channels 1 and 2 (blue and red curve).    Homozygous mutant blood samples, channel 1 (ROX) will show a strong signal and amplification plot (red curve) and channel 2 (Cy5) none or a very low signal and amplification plot (blue curve). So the signals are reversed on homozygous mutant samples.   

DirectBlood Genotyping Kit performance with a hybridization probe assay

Here we are detecting the same SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hybridization probe assay.

Three blood samples with known genotype were used. All the genotypes can clearly be identified. The blue curve shows the specific wild type (homozygous G;G) peak at 61°C. The red curve shows the specific mutant (homozygous A;A) peak at 53°C. A heterozygous case (A;G) will result in both melting curve peaks, respectively at the same specific temperature for wild type and mutant (orange curve).

DirectBlood Genotyping PCR Kit is a novelty on the market. It allows genotyping in real-time and without any previous DNA isolation. You simply save time and money: directly use EDTA blood without any extraction steps.
The DirectBlood Genotyping PCR Kit is optimized to function with a wide range of different SNPs to detect possible nucleotide variations. It has been successfully tested with a variety of hydrolysis probes and hybridization probes.

t-Boc-N-amido-Tri-(propargyl-PEG10-ethoxymethyl)-methane is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The Boc group can be deprotected under mild acidic conditions to form the free amine. Reagent grade, for research use only.

t-Boc-N-amido-Tri-(propargyl-PEG10-ethoxymethyl)-methane is reactive with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The Boc group can be deprotected under mild acidic conditions to form the free amine. Reagent grade, for research use only.

OverView

The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.

The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).

Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.

Protocol

The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps before running RT-qPCR.

• gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
• Heat-labile dsDNase can easily be inactivated.
• This procedure minimizes pipetting steps and reduces hands-on time.
• The Heat&Run kit is especially well suited for high throughput experiments.

The high activity of the HL-dsDNase at low temperature and its heat-lability makes it ideal to use in a combination with a heat activated RT enzyme. The contaminating genomic DNA is cleaved at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.

Do you require gDNA removal in applications other than RT-qPCR? Contact our support team for assistance in implementing dsDNase treatment in your workflow.

Kit Contents

• 10X reaction Buffer
• HL-dsDNase (50 or 250 reactions)

The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.

The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).

Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.