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- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
低温
- 保质期:
详见说明
- 库存:
99
- 供应商:
上海圻明生物
- 规格:
250 mL
DNA级异硫氰酸胍溶液,5M上海圻明生物优势供应。更多产品资料欢迎免费咨询。
One of the many important uses of PCR technology is that it can be used to label DNA probes with high specific activity. PCR technology has high specificity, and can synthesize probe DNA fragments in quantities within 1~2h if [α-32P]dNTP or other markers are added to the substrate
dNTPs, the probe DNA can be well labeled during DNA synthesis, and the incorporation rate of the marker can be as high as 70%~80%. Therefore, PCR labeling technology is particularly suitable for large-scale detection and non-radiolabeling. The disadvantage of this method is that a specific pair of PCR primers is synthesized.
Labeling can also be achieved by using small fragments prepared from probe DNA as primers.
Solution preparation
1. Prepare a stock solution
Unless otherwise stated, all unused stock solutions should be divided into disposable aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.
1.1* Acid Stock Solution (125X):
Add 20 μL DMSO to *ate (component B) to make a 125X* acid stock solution.
2. Prepare standard solutions
*Salt standard solution
Add 50 μL of 1 mM KH2PO4 (Component C) to 950 μL of deionized water or enzyme reaction buffer to give a 50 μM * saline standard solution (PS7). A 50 μM * saline standard solution (PS7) was taken and serially diluted 1:2 to obtain a serially diluted phosphate standard with deionized water or enzyme reaction buffer.
3. Prepare a working solution
Add 20 μL of 125X* stock solution to 2.5 mL of sterile H2O and mix well to make a working solution of *salt. Avoid potential Pi contamination. Note: Avoid direct exposure of *salts (component B) to light. Due to the high sensitivity of this assay to Pi, it is extremely important to use Pi-free labware and reagents.
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文献和实验g/mL 无 DNA 酶的 RNA 酶,37 ℃ 温育 1 h,以除去残留的 RNA。6. 重复步骤 4、5。1 g 细胞大约能提取到 2 mg DNA。(二)从植物组织中提取基因组 DNA【实验试剂与器材】1. CTAB 抽提液:2% (w/v) CTAB (十六烷基三甲基溴化铵),100 mM Tris.Cl,pH8.0, 20 mM EDTA,pH8.0,1.4M NaCl2. CTAB/NaCl 溶液:10% CTAB,0.7M NaCl配制方法:在 80 mL 双蒸水中溶解 4.1 g
。 a.时间越长,DNA的产量越多,污染也越多 7. 12000转速离心5-10min,去上清,倒置于吸水纸上数分钟,加入700µl 70%的酒精,上下颠倒数次,洗涤沉淀。12000转离心5-10m,去上清,倒置于吸水纸上数分钟。重复一遍。 8. 12000转速离心5-10min,去上清,倒置于吸水纸上数分钟,最后置于超净工作台中吹干。 9. 在离心管中加入30-50µl的灭菌去离子水或者TE溶液,4℃放置数小时,使其充分溶解。 10
大量获取玉米叶片DNA为例的实验流程: (1)选取适量的玉米幼苗新鲜叶片,以纱布捆扎,做好标记,置于液氮中保存; (2)研钵以液氮预冷,将玉米叶片置于研钵中,倒入液氮,迅速研磨至粉末级,期间时时加入液氮,防止研磨过程中DNA被降解; (3)将研磨好的粉末移入灭菌后的50mL抽提试管中,用移液管加入10mL月桂酸钠抽提缓冲液,反复缓慢的摇匀; (4)用移液管向50mL抽提试管中加入等体积(10mL)酚:氯仿:异戊醇(25:24:1),反复缓慢的摇匀; (5)12000 rpm,4°C
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