12/21/2023 0 Comments Noti restriction enzymeCan the change in buffer preference of the HF enzyme be advantageous?.When should I choose the High Fidelity (HF ®) version of the enzyme?.Why does the HF version of the enzyme have a different recommended buffer than the wild type enzyme?.What does HF® refer to following the name of a restriction enzyme?.What is the difference between NotI-HF and NotI?.Is there a difference in cutting close to the ends between NotI-HF and NotI?.Is there any difference in the methylation sensitivity between NotI-HF and NotI?.coli strain that carries the cloned and modified NotI gene from Nocardia otitidis-caviarum (ATCC 14630) This product is related to the following categories: Restriction Endonucleases: N-O, High-Fidelity (HF®) Restriction Endonucleases Products, Time-Saver Qualified Restriction Enzymes Products This product can be used in the following applications: Fast Cloning: Accelerate your cloning workflows with reagents from NEB, Restriction Enzyme Digestion The resolution isĪt the single nucleotide level. Is determined by capillary electrophoresis and peak analysis. Restriction Enzyme Competitor Study: Nuclease ContaminationĮcoRI, NotI, and BamHI from multiple suppliers were tested in reactionsĬontaining a fluorescent labeled single stranded, double stranded blunt,ģ’overhang or 5’ overhang containing oligonucleotides. Of our HF restriction enzymes are shown below. Examples of nuclease contamination studies for some NEB extensively performs quality controls on all standard and high-fidelity Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 with the flexibility to digest overnight without degradation to DNA. HF enzymes also exhibit dramatically reduced star activity. Pustaka genom dengan sisipan fragmen ~40 Kb yang membawa gen MTGase telah berhasil dikonstruksi dengan menggunakan fosmid pCC1FOSTM di dalam E.coli EPI300-TIR.High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart Buffer single-buffer simplicity means more straightforward and streamlined sample processing. Berdasarkan sekuen gen 16S rRNA, isolat TTA 02 SDS 14 memiliki kekerabatan terdekat dengan Streptomyces thioleteus dengan homologi 99%. Sekuen parsial gen MTGase dari isolat tersebut memiliki homologi 93 % dengan MTGase dari Streptomyces cinnamoneus. Analisis molekuler dengan PCR membuktikan bahwa isolat tersebut mengandung gen MTGase. Hasil penelitian menunjukkan bahwa penapisan terhadap bakteri Streptomyces spp penghasil MTGase yang diisolasi dari berbagai jenis tanah di Indonesia, mendapatkan satu isolat yang potensial yaitu TTA 02 SDS 14. Untuk itu, penelitian ini dilakukan melalui beberapa tahapan yaitu, penapisan bakteri Streptomyces spp penghasil MTGase, identifikasi isolat penghasil MTGase dan pembuatan pustaka genom dengan mengkloning fragmen DNA 40 kb yang mengandung gen penyandi MTGase. Penelitian ini bertujuan untuk mendapatkan bakteri yang menghasilkan MTGase dan mengklon fragmen DNA penyandinya. Pendekatan teknologi rekayasa genetika sangat diperlukan untuk mendapatkan hasil yang lebih baik sehingga dapat dikembangkan dalam skala industri. Selain itu proses pemurnian cukup sulit karena memerlukan beberapa tahap pemurnian. Produksi enzim Microbial Transglutaminase (MTGase) dari strain liar mempunyai beberapa kelemahan yaitu selain pertumbuhan selnya lambat, produk yang dihasilkan sedikit dan protein yang dihasilkan juga bercampur dengan protein lainnya. 2.3.2.13) merupakan enzim yang mengkatalisis reaksi perpindahan gugus asil antara residu glutamin (Gln) yang berfungsi sebagai donor asil dan residu lisin (Lys) sebagai aseptor yang membentuk ikatan silang (crosslinking) ε-(γ-glutamyl) lisin isopeptida yang menghasilkan ikatan kovalen inter atau intramolekuler yang berikatan silang dengan protein makanan.
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