DNA 断片選別のための効率的なソリューションを提供する方法?

現在のところ, 分子生物学研究における多くの問題では、サイズに応じて DNA を分類する必要があります, これはフラグメントソートとして知られています. アガロースゲルでの電気泳動により、以下の範囲の DNA を分離できます。 10 ⁵～数百の塩基対. 対照的に, アクリルアミドゲルでの電気泳動により、数千塩基対の範囲の DNA を分離できます。. これらの分離プロセスで高分解能を達成するには, 2 つのフラグメントを分離するためのサイズの差は次のとおりです。 10% 最大の核酸含有量 5 に 10 ミリグラム.

選択的沈殿法, ポリエチレングリコールと生体磁気ビーズの組み合わせ, フラグメントソートに対するより効率的で便利なアプローチとして際立っています. 高濃度のPEGおよびNaCl溶液中, PEGはDNA分子の外側の水和層を破壊します, それらが凝集して沈殿する原因となる. This exposes the negatively charged phosphate groups, which then form a ‘salt bridge’ with the carboxyl groups on the magnetic beads’ surface, allowing DNA to be adsorbed onto the beads.

The longer the DNA, the more negatively charged phosphate groups are exposed on the surface, making the entire molecule more negatively charged and easier to adsorb onto magnetic beads, and it can be recovered with lower concentrations of PEG and NaCl. The shorter the DNA, the higher the concentration of PEG and NaCl needed to break down the hydration layer on its surface thoroughly, exposing enough negatively charged phosphate groups to be adsorbed by magnetic beads and then recovered. If you want to recover shorter DNA fragments, you need to add a larger volume of magnetic beads. The ratio of magnetic bead suspension to the sample also affects the size of fragments screening. As the increasing of the concentration of magnetic bead suspension, the magnetic beads will increase their adsorption capacity for small fragments in addition to large fragments.

When a specific length of DNA fragment is to be separated by magnetic beads, two rounds of sorting are typically performed. In the first round, the magnetic beads bind to DNA with higher molecular weight, which is then removed by discarding the beads. The second round involves the remaining product, where the magnetic beads bind to DNA with higher molecular weight and remove DNA with lower molecular weight by discarding the supernatant.

Both double-stranded DNA, single-stranded DNA, and RNA can dissociate negatively charged phosphate groups in solution so they can be purified and recovered with magnetic beads. しかし, single-stranded DNA and RNA cannot be screened for fragment size. The secondary structure of double-stranded DNA is generally a relatively stable linear double-helix molecule. In the presence of PEG, the number of negatively charged phosphate groups dissociated is positively correlated with its length. しかし, RNA and single-stranded DNA have complex secondary structures, which prevent the exposed negative groups from being positively correlated with length, making it impossible to screen fragments like DNA. 加えて, due to the varying stability of DNA and RNA at different pH values, the pH of the buffers used for purifying DNA and RNA are different because DNA and RNA have different stability under different pH conditions.

Shanghai Lingjun Biotechnology fragment sorting magnetic beads are independently developed and produced from raw materials to finished magnetic beads, with strict quality control to ensure that the performance and consistency of the final product meet high standards and ensure a stable supply of the product. The company continuously invests in research and development, always maintaining an absolute emphasis on product quality, customer satisfaction, and market competitiveness.

サプライヤー紹介

上海霊軍生物技術有限公司, 株式会社に設立されました 2016 生体磁性材料と核酸抽出試薬の専門メーカーです.

核酸抽出・精製の経験が豊富です。, タンパク質の精製, 細胞分離, 化学発光およびその他の技術分野.

当社の製品はさまざまな分野で広く使用されています, 医療検査など, 遺伝子検査, 大学の研究, 遺伝子育種, 等々. 製品のご提供だけでなくOEMも承ります, ODM, その他のニーズ. 関連するニーズがある場合, お気軽にお問い合わせください。 sales01@lingjunbio.com.