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Nucleic Acid

GRiSP offers one of the most innovative and high performance options on the market for directPCR applications - Xpert directXtract PCR Kit.

One of the main applications for this kit (despite not limited to) is Mouse Genotyping. Not only it's an application with intensive reagent numbers consumption, but also one where time is highly relevant.

mouse genotyping

 

The workflow, show in the flyer on the left, contains three steps, and a total time of only 90 minutes.
It consists on the combination of GE60 (Xpert directXtract PCR Kit) and GB01 (SGTB DNA Electrophoresis Buffer).

 grisp kit

Replace Ice Bucket

 
Traditional disadvantages:
1. Not easy to organize samples. Tubes can’t stand straightly on ice
2. Ice never stops melting, and sample is easy to be contaminated when soaking in melting water
3. Ice-maker machine wastes too much energy because it has to operate every minute to maintain ice making
 
eCooler’s advantages:
1. Easy to organize samples, with different composition of rack
2. Stable temperature, never melt and keep samples clean
3. Save energy, diminishes 2.67MT/year CO2 emission
 

 
Replace other cooling systems, ex. dry bath
 
Other cooling system’s disadvantages:
1. Very slow cooling speed, usually costs more than 30 minutes from room temperature to 4 ℃
2. Only can stay in place with electricity plug hole, and can’t be moved around
3. Some cooling boxes have to be stored in refrigerator before using. Although they needn’t electricity, they occupy a lot of spaces in refrigerator and can’t make sure stable temperature after taken out
 
eCooler’s advantages:
1. The fastest cooling speed in the world, 10 minutes from room temperature to 4 ℃
2. Portable, temperature maintained in 4 ℃ within 30 minutes
3. No need to occupy refrigerator’s space, and promise stable temperature in ± 0.5 ℃
 
 
3. Combined with PCR
 
Why does PCR need eCooler?
No matter general PCR reaction or real-time PCR reaction, all reagents had better to be mixed in low temperature. dNTP and Polymerase may start wrong reaction under improper temperature, before putting tube into PCR machine. Researchers have to keep them inactive. After PCR reaction, researchers still need to keep PCR products under low temperature for following experiment.
 
eCooler is convenient for researchers to prepare PCR reaction solution at low temperature!
 

4. Easy to Freeze Cell
 
How eCooler helps cell-freezing?
When freezing cell, researchers have to centrifuge down cells from medium, and add freezing media to suspend. Suspending have to be operated on ice for protecting cell and them adding DMSO. Traditionally, researchers have to operate on ice, but cell is very sensitive to contamination and unstable temperature. A little mistake such as drop a piece of ice into tube could result in horrible consequences. Especially in P2/P3 cell culture room, this kind of laboratory has to stay isolation from out-world. So most of them can’t install ice-maker machine.  When researchers have to use ice, they have to bring ice from out-world. It needs a complex sterilize process. Put eCooler in P2/P3 cell culture room could solve this problem. It doesn’t need to connect water pipe, doesn’t need to bring from outside, and is easy to move to liquid nitrogen barrel for next freezing step.
 

Improved management of hepatitis D

fusion spectra

 

HDV is a small, defective RNA virus that can infect only individuals who have HBV; worldwide
more than 15 million people are co-infected. HDV increases the severity of chronic HBV infection,
frequently leading to cirrhosis, hepatic decompensation or hepatocellular carcinoma. The prevalence
of HDV is declining in some endemic areas but increasing in northern and central Europe
because of immigration. Interferon-alpha is currently the only available treatment option leading
to suppression of HDV RNA in 25 % - 30 % of patients [

pdfImproved management of hepatitis D

Amp_Tec_logoExpressArt® mRNA amplification technology for intact RNA: How does it work?

 

Briefly, ExpressArt ® technology for Standard kits is based on the following steps: Conversion of mRNA to cDNA is achieved with an anchored oligo(dT) primer in the first reverse transcription reaction. No promoter sequence is present in the primary cDNAs. Then, all RNAs (including mRNAs and rRNAs) are digested with a mixture of heat-labile RNases. Single stranded cDNAs are converted to double stranded DNAs with a special primer construct, the TRinucleotide primer (BOX-random-3'-trinucleotide). This 30-mer primer contains a unique 21-mer BOX, followed by six random nucleotides, and a trinucleotide sequence at the 3'-end. We use a mix of several primers with different 3'-terminal trinucleotides.

The trinucleotides determine potential primer elongation sites, that are discontinuously distributed over annealed templates. Consequently, preferential priming near the 3'-end of the template occurs. To minimise primerderived artefacts, the T7-promotor sequence is introduced only in the 2nd synthesis of dsDNA. This double stranded cDNA is the template for the first amplification by in vitro transcription. All amplified RNAs contain the same 3'-terminal 21-mer BOX sequence. For second and third amplification rounds, full-length cDNAs are obtained using this 21-mer BOX sequence as primer. RNA removal (with heat-labile RNases) is followed by second strand cDNA synthesis, introducing a T7-promotor sequence in double stranded cDNA templates for second (or third) amplification by in vitro transcription.

 

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