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- RPN303B
- 2025年07月12日
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大量
- 规格：
30cmx3m
带正电荷的尼龙膜与核酸高达600μg/cm2的结合能力。建议使用放射性和非放射性的化学发光法和chemifluorescence检测系统。
Amersham公司Hybond™- N +产品在传统的Southern杂交有着出色的灵敏度。核酸样品可能是固定的，通过紫外线照射或烘烤，虽然紫外线固定是最好的最大的重复性。 Hybond N +是不适合碱印迹和固定。
Amersham Hybond™-N+
- Positively charged nylon membrane with a binding capacity for nucleic acids up to 600 µg/cm2¾recommended for use with radioactive or nonradioactive chemiluminescence and chemifluorescence detection systems.
- Recommended for all high density membrane arrays.
Technical Information
Amersham Hybond™-N+ yields excellent sensitivity in conventional Southern blotting. Nucleic acid samples may be fixed by UV exposure or baking, although UV fixation is best for maximum reproducibility. Hybond-N+ is not suitable for Alkali blotting and fixation.
- Multiple reprobing is possible without loss of membrane integrity.
- QC tested in genomic Southern blots using GE Healthcare products.
- Special sizes available on request.
Human genomic DNA (1, 0.5, 0.125, 0.06 and 0.03 µg loadings) digested with HindIII and transferred to Hybond-N+. Hybridizaton with A. N-ras, B. p53, and C. bcl-2 labeled DNA probes and detected using Gene Images™ random-prime labeling with CDP-Star™ detection.
Membranes should be stored at room temperature in a clean dry atmosphere away from excessive heat, light, and noxious fumes. To prevent contamination, handle membranes using gloves or blunt-ended forceps. Hybond-N+ is stable for up to 3 years. Membranes should be kept in the bags in which they are received. Performance is consistent when stored under the recommended conditions.
| TECHNICAL SPECIFICATIONS | |
| Pore size | 0.45 µm |
| Nucleic acid size required | > 50 bp |
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文献和实验，然后加入戊二醛，在戊二醛的化学交联作用下与带负电荷的单链DNA共价结合，从而标记DNA。标记的探针DNA与膜上的单链DNA杂交形成双链DNA，探针上的HRP在H2O2存在下，催化H2O2还原，同时使鲁米诺氧化并伴随蓝光产生，在增强剂的作用下使产生的光加强，从而可在X光片上检测。 首先将经限制性内切酶酶解的DNA片段，经琼脂糖凝胶电泳分离以及在凝胶上经NaOH处理使之变性，然后将尼龙膜(Hybond-N+)放在凝胶上，使之按原有顺序将条带转移至膜上并固定起来，这是Southern转膜过程
ml DEPC H2O~1.8% dye. Mix 95 uL Formamide with 5 uL 1.8% Bromophenolblue/Xylene cyanolFF Note: can use less dye. Hybond N+: Amersham RPN303B ULTRAhyb-Oligo Hybridization Buffer: Ambion cat # 8663, keep in refrigerator T
Screening BAC filters with non-radioactive probes
(Protocol for a single high-density BAC colony filter (HDR filter) of 22 cmx22 cm) to be screened using Amersham''s ECL hybridization kit; this protocol was kindly contributed by Dr. Kazuei Mita from the National Institute of Agrobiological
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