Highlights
- Rapid, simple, high-throughput (96-well) method for DNA isolation from tough-to-lyse plant and seed samples.
- Ultra-high density BashingBeads are fracture resistant and chemically inert.
- Zymo-Spin technology coupled with filtration removes polyphenolic PCR inhibitors from the DNA product.
Description
| Applicable For | All sensitive downstream applications such as qPCR and Next-Generation sequencing. |
|---|---|
| Elution Volume | ≥ 50 µl |
| Equipment | Centrifuge w/ microplate carriers, 96-well plate/block disruptor or pulverizer. |
| Processing Volume | ≤ 80 mg |
| Purity | A260/A280 nm ≥1.8. |
| Sample Source | Leaves, stems, buds, flowers, fruit, seeds, etc. |
| Sample Storage | DNA stored at ≤ -20°C. |
| Size Range | Capable of recovering genomic DNA up to and above 40 kb. Typical fragment sizes range from 25 kb - 35 kb. If present, parasitic and viral DNA will also be recovered. |
| Type | Total DNA |
| Yield | ≤ 5 µg total DNA |
Q1: Can you provide a list of the tested plant species?
We currently do not have a list of plants, aside from what we have shown in the protocols:–A.thaliana–Juniper–Milkweed Leaf–Milkweed Leaflet–Milkweed Pre-Flowering Bud–Corn Kernel–Sunflower Seed–Nicotiana sp.
Q2: My lysate seems viscous. What is causing this to happen? How can I fix this?
A viscous sample can indicate incomplete sample lysis. Try using less of your sample and optimize bead beating conditions (duration, speed, time) to ensure samples are thoroughly lysed. After bead beating, pellet the cell debris before moving on. Adding more Genomic Lysis buffer to the lysate can help dilute and deproteinate the sample, making the sample less viscous and more suitable for DNA recovery.
Q3: What is the purpose of Silicon-A-HRC Plate step?
Environmental samples often contain inhibitors such as polyphenolics, humic/fulvic acids, tannins, melanins, etc. that often co-purify and affect downstream applications such as PCR. The Zymo-Spin III-HRC column removes these polyphenolic PCR inhibitors to recover DNA that is ready for sensitive downstream applications such as Next-Gen Sequencing, quantitative PCR, etc.
Q4: Is it necessary to add beta-mercaptoethanol? Can this step be substituted or omitted?
Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM). However, if bead beating is optimized and lysis is efficient, the addition of BME is not necessary and can be omitted.
Q5: When can an RNase A treatment be implemented in the protocol?
The Quick-DNA kits recover RNA-free genomic DNA. The selective chemistry allows for binding of double stranded DNA to the column and for RNA to flow through. No RNase A treatment is required when processing samples within kit capacity.
| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| S6002-96-2 | ZR-96 BashingBead Lysis Rack | 2 mm | $192.00 | |
| C2009 | Silicon-A-HRC Plate | 2 Plates | $405.00 | |
| C2003 | Elution Plate | 2 Plates | $19.00 | |
| C2002 | Collection Plate | 2 Plates | $22.00 | |
| C2007-4 | 96-Well Plate Cover Foil | 4 Foils | $10.00 | |
| P1001-2 | 96-Well Block | 2 Blocks | $18.00 | |
| C2001 | Silicon-A Plate | 2 Plates | $129.00 | |
| D6035-1-30 | Prep Solution | 30 ml | $18.00 | |
| D6001-3-40 | BashingBead Buffer | 40 ml | $29.00 | |
| D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
| D3004-2-100 | g-DNA Wash Buffer | 100 ml | $30.00 | |
| D3004-1-150 | Genomic Lysis Buffer | 150 ml | $73.00 |
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,表示体内感染了HBV,因而是一种特异性标志。HBsAg阳性见于:①急性乙型肝炎的潜伏期或急性期(大多短期阳性);②HBV致的慢性肝病、迁延性和慢性活动性肝炎、肝炎后肝硬化或原发性肝癌等。③无症状携带者。
(2)抗HBs:表示曾感染过HBV,均已得到恢复,并且对HBV有一定的免疫力。
(3)HBcAg与抗HBc:由于
HBcAg主要存在于肝细胞核内,并仅存在于Dane颗粒中。因此,对病人血清不能检测HBcAg,而测抗HBc。血清内抗HBc阳性反映:①新近有过HBV感染;②体内有HBV增殖;③有助于诊断乙型肝炎。
(2)HBcAg和抗HBe:HBcAg的存在常表示病人血液有感染性。
HBcAg阳性揭示病人肝脏可能有慢性 损害,对预后判断有一定帮助。抗HBe阳性对病人可能有一定的保护力。
检测乙肝抗原与抗体的实际用途:
(1)筛选供血员。
(2)可作为乙肝病人或携带者的特异性诊断。
(3)对乙肝病人预后和转归提供参考。
(4)研究乙肝的流行病学,了解各地人群对乙肝的感染情况。
(5)判断人群对乙肝的免疫水平。
抗原与其所诱导产生的抗体或致敏淋巴细胞特异性结合的能力。抗原性的强弱与抗原分子的大小、化学成分、抗原决定簇的结构、抗原与被免疫动物亲缘关系的远近等有密切关系。通常认为抗原的分子量愈大、化学组成愈复杂、立体结构愈完整以及与被免疫动物的亲缘关系愈远,则抗原性愈强。抗原的物理状态也对抗原性发生影响,例如蛋白质,聚合状态的比单体的抗原性强,一般球形分子的比纤维形分子的抗原性强。抗原加入佐剂改变物理状态后,抗原性也得到增强。例如,分子量高达10万的明胶由于缺乏苯环氨基酸,稳定性较差,在进入机体后容易被酶降解成低分子物质,如果加入少量酪氨酸(苯环氨基酸),就能增强其抗原性。
中文名:抗原性
外文名:antigenicity
别称:免疫反应性
抗原性介绍:流感病毒按抗原性

