Hieff Unicon ™ ColorGPS QPCR Master Mix (ROX Tinggi) _ 11190es

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Hieff Unicon ™ ColorGPS QPCR Master Mix (ROX Tinggi) _ 11190es

Hieff UNICON™ ColorGPS qPCR Master Mix (High Rox) merupakan pra-solusi untuk amplifikasi PCR kuantitatif 2xreal-time, yang memiliki intensitas fluoresensi tinggi, sensitivitas dan spesifisitas tinggi, serta hasil amplifikasi tinggi. Komponen inti, Hieff UNICON™ Taq DNA Polymerase, diaktifkan secara panas melalui metode antibodi, yang secara efektif dapat menghambat amplifikasi non-spesifik yang disebabkan oleh penempelan primer selama persiapan sampel. Pada saat yang sama, formula tersebut menambahkan faktor untuk meningkatkan efisiensi amplifikasi reaksi PCR dan fasilitator untuk menyamakan amplifikasi gen dengan kandungan GC yang berbeda (30~70%), sehingga PCR kuantitatif dapat memperoleh hubungan linier yang...

Frequently Bought Together

Fitur

Pelacakan Pipet: Mengurangi Risiko Kesalahan Penambahan/Kehilangan

Amplifikasi Luar Biasa: Linearitas yang baik pada rentang linier yang lebar

Presisi Tinggi: Dapat secara akurat membedakan templat dengan perbedaan konsentrasi 2 kali lipat

Pengulangan yang Baik: Variabilitas antar sumur kecil dalam sumur replikasi, menghasilkan pengulangan yang lebih baik

Stabil dan Handal: Sistem reaksi dapat disimpan pada suhu ruangan selama 24 jam, dan produk tetap tidak terpengaruh setelah 20 siklus beku-cair

Aplikasi

Analisis Perbedaan Ekspresi Gen

Analisis Kuantitatif Absolut

Spesifikasi

Konsentrasi	2 Bahasa Indonesia:(Campuran qPCR)
Metode deteksi	Metode pewarnaan
Metode PCR	qPCR
Polimerase	Taq DNA Polimerase
Jenis sampel	DNA
Peralatan aplikasi	Biosystems 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne, StepOnePlus dan instrumen PCR kuantitatif fluoresensi lainnya yang memerlukan penambahan konsentrasi tinggi Pewarna Referensi ROX
Tipe Produk	Premix untuk PCR kuantitatif fluoresensi waktu nyata
Lamar ke (lamaran)	Ekspresi Gen

Komponen

Komponen No.	Nama	1tahun 1190Bahasa Indonesia: ES03	1tahun 1190Bahasa Inggris ES08	1tahun 1190Bahasa Indonesia: ES60
11190-A	UNICON yang kuat™ qPCR GPS warna Campuran Master (Tinggi (Kotak)	1 ml air	5×1ml	100x1ml
11190-B	Penyangga Pengenceran 10×	1 ml air	1 ml air	20×1ml

Pengiriman dan Penyimpanan

Produk ini harus disimpan pada suhu -25~-15℃ selama 1 tahun.

Angka

Gambar 1. Hasil yang dapat diandalkan dapat diperoleh dalam rentang dinamis.

Menggunakan 2 µL templat plasmid dengan gradien 10^1-10^7, gen terkait diperkuat. Hieff UNICON™ Campuran utama ColorGPS dapat secara efektif mendeteksi rentang jumlah templat sebanyak 7 kali lipat, mencapai linearitas yang baik dalam rentang linear yang luas.

Angka2Dapat secara akurat membedakan perbedaan konsentrasi templat sebesar 2 kali lipat.

Menggunakan 2 µL plasmid yang diencerkan dalam gradien 2 kali lipat sebagai cetakan, gen terkait diperkuat. Hieff UNICON™ Campuran utama ColorGPS dapat secara akurat mengatasi perbedaan konsentrasi templat.

Angka3Variabilitas antar sumur kecil diamati, menunjukkan pengulangan yang lebih baik.

Dengan menggunakan lusinan salinan plasmid sebagai templat, amplifikasi dilakukan dengan berbagai reagen kuantitatif fluoresensi sesuai dengan prosedur yang ditetapkan dalam buku petunjuk masing-masing. Nilai SD menunjukkan bahwa, dibandingkan dengan reagen transkripsi balik lainnya, produk baru ini memiliki perbedaan yang lebih kecil antara sumur replikasi dan pengulangan yang lebih baik.

Angka4Lebih fleksibel, cocok untuk prosedur standar atau cepat.

Dalam prosedur standar, waktu denaturasi dan waktu ekstensi ditetapkan masing-masing pada 10 detik dan 30 detik, sedangkan dalam prosedur cepat, waktu denaturasi dan waktu ekstensi ditetapkan masing-masing pada 3 detik dan 10 detik untuk mendeteksi nilai Ct yang sesuai dengan 10 gen target.Hasilnya menunjukkan bahwa nilai Ct yang diperoleh dengan produk baru pada prosedur standar dan cepat memiliki korelasi yang kuat, dan produk baru dapat mencapai hasil yang baik menggunakan prosedur cepat.

Angka5Setelah sistem reaksi disiapkan, sistem dapat tetap stabil pada suhu ruangan selama 24 jam.

Menggunakan berbagai gen dari tikus, sel 293 manusia, dan Arabidopsis thaliana sebagai target, setelah menyiapkan sistem reaksi, sistem tersebut ditempatkan dalam gelap pada suhu yang berbeda selama 24 jam sebelum dideteksi. Dibandingkan dengan pemrosesan langsung, perbedaan deteksi untuk berbagai gen tidak melebihi 0,5 Ct, yang menunjukkan bahwa hasil yang stabil juga dapat diperoleh dengan operasi berdurasi panjang.

Dokumen:

Buku Panduan

1tahun 1190_Pedoman_Versi EN20241218Bahasa Indonesia: .pdf

Storage

This product should be stored at -25~-15℃ for 18 months.

FAQ

How can one select the appropriate fluorescent quantitative reagents based on different instrument models?

Please refer to the "Yeasen Biotechnology Dye Method Fluorescence Quantitative PCR Mix Selection Table" for details. Click here to learn more and help you choose the appropriate reagents.

How should the reagents be mixed before use?

For molecular enzyme reagents, simply invert and mix them together, then perform a gentle centrifugation. It is not recommended to vigorously shake to mix them.

How to use the yellow diluent? Can it be omitted?

Refer to the manual. Ultimately, it is necessary to ensure that after being mixed with the template, the volume is exactly 1×.
For example, for 20 μL of cDNA stock solution, add 180 μL of sterile ultrapure water to dilute it 10 times, resulting in 200 μL of diluted template. Take 90 μL of the diluted template and add 10 μL of 10× Dilution Buffer, mix well, and then use it. You can choose not to do this, but it will not affect the result.

Do the cDNA templates need to be diluted? And by what factor?

You can choose to use the cDNA stock solution or dilute it. Just make sure that the Ct value of the gene you want falls within the range of 15 to 30. Generally, for every 10-fold dilution of cDNA, the Ct value increases by 3.3.

The reaction system has been prepared, but it can't be loaded onto the machine. What should we do?

Store in a dark place and the reaction system must not be left open. These two points are essential. It can be placed in a 4℃ refrigerator or at room temperature for 24 hours. Before using, gently shake and centrifuge the sample before loading it onto the instrument. Before loading, carefully check if the tubes or plates have been contaminated.

The data report on the instrument shows a "BadRox" error. What should I do?

The possible reasons are as follows:
1)The instrument has not been calibrated for a long time or it is aging. Usually, the instrument needs to be calibrated once a year. It is necessary to have an instrument engineer perform the calibration or replace the aging components.
2)A 10 μL reaction system was used. The normal system is a 20 μL system. Using half of the normal system would result in an excessively low total concentration of Rox, increasing the probability of BadRox occurrence. It is recommended to conduct the experiment using the 20 μL system.

Can the qPCR products of this product be run on a gel electrophoresis?

The specificity of qPCR products can usually be determined through the melting curve. If it is still necessary to determine the products through gel electrophoresis, the qPCR product can undergo electrophoresis. Electrophoresis requires Loading Buffer, and the product cannot be directly loaded for electrophoresis.

After the amplification process is completed, no amplification curve was observed?

1)Confirm whether the signal acquisition function is enabled in the program. If a two-step method is used, the signal acquisition should be set during the annealing extension stage.
2)Is the length of the product too long? If the length of the product is set to 500bp or more, this situation may occur. It is recommended that during primer design, the length of the product be controlled within the range of 80-300bp.
3)Are the primers inappropriate? If the primers are not designed correctly, they need to be redesigned. If the primers are correctly designed but seem to be degraded, PAGE electrophoresis can be used to check the integrity of the primers.
4)Template degradation: Prepare the template anew and repeat the experiment.

If the NTC shows a CT value, what should we do?

The analysis is conducted by observing the melting curve.
The melting curve shows that the melting peak shape of NTC does not overlap with that of the corresponding gene, and the TM value of NTC is smaller than that of the corresponding gene. In this case, the product corresponding to NTC is a primer dimer. If the melting curve peak shape of the corresponding gene is a single peak, the collection of the signal of the related gene will not be affected.
The melting curve shows that the melting peak shape of NTC overlaps with that of the corresponding gene. In this case, the product corresponding to NTC is likely to be the product of the corresponding gene. The system has been contaminated. We need to check one by one whether the water, primers, or reagents have been contaminated. If none of them have been contaminated, it might be aerosol contamination. Based on the obtained amplification data, the ΔCT value is calculated to determine whether the contamination has a significant impact on the system. If ΔCT ≥ 5, it indicates that the contamination has a minor effect and can be ignored.

What should we do when the CT values are shown for NRT?

It indicates the presence of genomic DNA contamination. It is recommended to use an RNA extraction kit that removes the genome for RNA extraction or during reverse transcription by adding a step to remove the genome. Alternatively, when designing the primers, a design across introns should be adopted.

If the CT value is less than 30, but the repeatability is very poor, what should we do?

Improve the accuracy of sample addition. Avoid adding small volumes and reduce the error of addition. This can be achieved by preparing a large sample system, for example, mixing primers, water, and qPCR mix into a large system or using the method of mixing primers and water into a large system. The large system needs to be mixed evenly.
Improve the accuracy of the pipette's aspiration. Pipettes usually need to be calibrated regularly, once a year.

What if CT is ≥ 30 but the repeatability is very poor?

At this point, due to the low abundance of the effective template, the randomness of collisions between the template and the primers increases, thereby causing a greater difference between the replicons. It is recommended to try adding 2-3 replicons, and then eliminate the data with large differences for data analysis.

What should we do if there are overlapping peaks in the melting curve?

The overlapping peaks in the melting curve are located to the left of the main peak. This might be a situation caused by primer dimerization. One possible solution could be to increase the annealing temperature, reduce the primer concentration, or redesign the primers.
The overlapping peaks in the melting curve are located to the right of the main peak.
1). It is possible that the non-specific amplification occurred due to poor specificity of the primers. The primers need to be redesigned.
2). It is possible that the template DNA was contaminated with genomic DNA. It is recommended to use an RNA extraction kit that removes the genomic DNA for RNA extraction, or to add a step of removing the genomic DNA during reverse transcription to reprepare the cDNA template.

Citations

Documents

Manual
11190ES-Hieff_UNICON_ColorGPS_qPCR_Master_Mix_High_Rox_-Ver.EN20241218.pdf

Brochure
qPCR_solutions.pdf

Applications

FAQ

Legal dan penafian

Produk ini hanya untuk keperluan penelitian dan tidak ditujukan untuk penggunaan terapeutik atau diagnostik pada manusia atau hewan. Produk dan konten dilindungi oleh paten, merek dagang, dan hak cipta milik Yeasen Bioteknologi. Simbol merek dagang menunjukkan negara asal, belum tentu pendaftaran di semua wilayah.

Aplikasi tertentu mungkin memerlukan hak kekayaan intelektual pihak ketiga tambahan.

Yeasen didedikasikan untuk ilmu etika, meyakini penelitian kami harus menjawab pertanyaan kritis sambil memastikan standar keselamatan dan etika.

Frequently Bought Together

Hieff Unicon ™ ColorGPS QPCR Master Mix (ROX Tinggi) _ 11190es

Dokumen:

Storage

FAQ

Citations

Documents

Applications

Hieff Unicon ™ ColorGPS QPCR Master Mix (ROX Tinggi) _ 11190es

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