Researchers often encounter problems with limited or insufficient DNA samples in downstream analyses. QIAGEN's Whole Genome Amplification Technology solves this problem by amplifying genomic DNA with small samples or precious samples, including single cells. The REPLI-g Kit accurately replicates raw DNA samples without bias, and the amplified DNA can be directly applied to a wide range of genetic analyses. What is whole gemome amplification? PCR-based WGA technology There are two main types of PCR-based WGA techniques: degenerate oligonucleotide PCR (DOP-PCR) technology (1) and pre-amplification primer extension (PEP) technology (2). The main difference between the two technologies is that the PEP technique uses random primers and low PCR annealing temperatures, while the DOP-PCR technique uses semi-degenerate oligonucleotides (such as CGACTCGAGNNNNNNATGTGG) and higher annealing temperatures. Taq DNA polymerase was used in both methods, limiting the length of amplification to 3 kb (average fragment length of 400–500 kb) and introducing errors into the amplified sequence. Moreover, studies have found that the genomic coverage of these techniques is not complete enough and will be biased in the amplification - due to the preferential binding of primers to certain specific regions, some sequences in the DNA amplification products are relatively increased. Multiple displacement amplification of WGA technology REPLI-g uses a thermostatic genomic amplification technique called multiple displacement amplification (MDA), which involves the binding of random hexamers to denatured DNA and strand displacement using Phi 29 polymerase at constant temperature. Synthesis. After the primers are added to each of the replacement strands, a branched DNA structure network is formed. Phi 29 polymerase does not dissociate from the genomic DNA template, which allows the resulting unbiased DNA fragments to be extended to 100 kb. The enzyme also has an exonuclease-correcting activity from the 3' end to the 5' end, which provides up to 1000-fold fidelity compared to Taq DNA polymerase-based methods (see PCR-based WGA technique above). Phi 29 polymerase is supported by a unique REPLI-g optimized buffer, which can easily overcome secondary structural difficulties such as hairpin loops, avoiding slippage, customization and polymerase dissociation during amplification. Such unbiased DNA fragments can be synthesized to a length of 100 kb. Advantages of MDA technology compared to PCR-based WGA method Traditional genomic DNA amplification methods include the time-consuming EBV transformed cell line establishment process, followed by PCR whole genome amplification steps using random or degenerate oligonucleotide primers. PCR-based WGA methods commonly used by other manufacturers also produce non-specific amplification interference and incomplete site coverage. DNA fragments shorter than 1 kb in length may be generated in some cases and are not available in many subsequent applications. In general, due to the use of low-fidelity Taq DNA polymerase, the resulting DNA contains a high rate of base mutation, thus forming an error-prone amplification, resulting in a single base pair mutation, STR. The occurrence of adverse consequences such as shortening and expansion. REPLI-g does not have these shortcomings. It can perform a highly uniform amplification of the entire genome, with minimal deviations and mutation rates during the reaction. The application of WGA technology using REPLI-g amplified genomic DNA is suitable for a wide variety of subsequent applications, including: The above information is from QIAGEN: http:// Specific product information: Linkage catalog number product name Linkage catalog price Y5-150023 REPLI-g Mini Kit (25) ¥ 2820 Y5-150025 REPLI-g Mini Kit (100) ¥ 8630 Y5-150033 REPLI-g UltraFast Mini Kit (25) ¥ 3100 Y5-150035 REPLI-g UltraFast Mini Kit (100) ¥ 9500 Y5-150043 REPLI-g Midi Kit (25) ¥ 7530 Y5-150045 REPLI-g Midi Kit (100) ¥ 25320 Y5-150052 REPLI-g Cell WGA & WTA Kit (12) ¥ 9850 Y5-150054 REPLI-g Cell WGA & WTA Kit (48) ¥ 33100 Y5-150063 REPLI-g WTA Single Cell Kit ¥ 14230 Y5-150065 REPLI-g WTA Single Cell Kit ¥ 45080 Y5-150090 REPLI-g Human Control Kit (25) ¥ 488 Y5-150126 REPLI-g Screening Kit (200) ¥ 17930 Y5-150243 REPLI-g FFPE Kit (25) ¥ 8050 Y5-150245 REPLI-g FFPE Kit (100) ¥ 23890 Y5-150343 REPLI-g sc Polymerase, Buffers, and Reagents ¥ 6810 Y5-150345 REPLI-g Single Cell Kit (96) ¥ 23070 Y5-151023 REPLI-g Mitochondrial DNA Kit (25) ¥ 3580 For more information about related products, please visit QIAGEN-Zhejiang and Jiangsu distributors. ; Read the original text: http:// Split Drape,U Split Drape,U Shape Drape,Surgical Split Drape Xinxiang Huaxi Sanitary Materials Co., Ltd. , https://www.huaximedical.com
As a method to increase the amount of limited DNA, genome-wide amplification technology emerged in 1992, which is particularly suitable for forensic identification and genetic disease research, as well as new technologies such as second-generation sequencing technology and CGH array (comparative genomic hybridization). Technical applications, the main problem of the latter is the limited number of DNA samples, but the analytical demand is considerable. A variety of WGA technologies have been developed in the industry, and the experimental scheme and replication accuracy between them are different.
WGA Technology - Comparison of PCR Technology and Multiple Displacement Amplification (MDA) Technology
The industry has developed a variety of WGA technologies; however, these technologies differ in their accuracy and ease of use. QIAGEN's REPLI-g Kit uses WGA technology called multiple displacement amplification to provide accurate, whole-genome amplification without bias.