Highlights
- Easy purification of high-quality DNA from whole blood, buffy coat, swabs, or cultured cells.
- Protocol excludes the use of Proteinase K and organic denaturants for biofluid and cell samples.
- Eluted, inhibitor-free DNA is ideal for PCR, endonuclease digestion, bisulfite conversion/methylation detection, sequencing, genotyping, etc.
Description
Elution Volume | ≥ 25 µl |
---|---|
Equipment | Microcentrifuge, vortex, centrifuge w/ microplate carriers |
Purity | High-quality DNA is eluted with DNA Elution Buffer or water. DNA is especially well suited for PCR and other downstream applications. A260/A280>1.8 |
Sample Source | Whole blood, plasma, serumcultured cells, buccal cells, tissue already digested with Proteinase K or mechanically homogenized from human, mouse, rat, etc. are effectively processed using this kit. |
Size Range | Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered |
Workflow | Unique lysis buffer system omits the need for Proteinase K digestion for biological fluids and cell culture samples. |
Yield | Up to 5 µg/well total DNA is eluted into ≥30 µl DNA Elution Buffer or water. Human whole blood yields 3-7 µg DNA per 100 µl blood sampled. Mammalian tissues already homogenized will yield 1-3 µg DNA per mg. |
Q1: What is the difference between Quick-DNA and Quick-DNA Plus kits?
The Quick-DNA is optimized for cells, soft tissues, and homogenized/digested samples using a single lysis/binding buffer. The Quick-DNA Plus kits contain an optimized Proteinase K for processing a wider variety of sample inputs, such as cells, blood, tissues, etc. The upgraded Quick-DNA Plus typically recovers more DNA.
Q2: I’m seeing some yield inconsistencies with my blood samples, what’s happening?
White blood cells, which are the major source of genomic DNA in blood, easily and quickly settles. Mix the blood sample well prior to taking an aliquot for purification.
Q3: Can the Quick-DNA kit be used with bacterial samples?
E. coli cells are easy-to-lyse and can be processed directly. For other microbes, additional pretreatment (e.g. enzymatic digestion or mechanical lysis) can be implemented and then processed with the Quick-DNA Kit. Alternatively, for an all-inclusive kit to process all microbes, use any of Zymo Research’s Environmental Kits (e.g. Quick-DNA Fungal/Bacterial, Quick-DNA Fecal/Soil, ZymoBIOMICS DNA, etc.) for DNA isolation.
Q4: Can I use the Quick-DNA kit to clean-up previously isolated DNA?
No, the kit is designed for direct use with biological samples. For clean-up of isolated DNA, please use the Genomic DNA Clean & Concentrator or the DNA Clean & Concentrator kits.
Q5: Can Quick-DNA process crude lysates?
Yes, add 4 volumes of Genomic Lysis Buffer to 1 volume of crude lysate, homogenized, or digested sample (see Cell Suspensions and Proteinase K Digested Samples) and proceed with the remainder of the protocol.
Q6: What is the purpose of adding beta-mercaptoethanol? Can this step be substituted or omitted?
Beta-mercaptoethanol is a reducing agent that helps break down proteins and improves DNA recovery and purity. Addition of beta-mercaptoethanol is recommended to enhance sample lysis, but can be substituted with dithiothreitol (DTT, final concentration of 10 mM) or omitted.
Q7: Is it possible to add an RNase A treatment to 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 specifications.
Q8: What are the expected yields for each sample type?
Keep in mind that there is sample to sample variability.
Sample Type | Input Amount | Expected Yield |
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Eukaryotic Cells | 1x106 Cells | 5-6 µg |
Skeletal, Heart, Lung, Brain Tissue | 1 mg | 1-3 µg |
Liver and Kidney Tissue | 1 mg | 3-5 µg |
Human Whole Blood | 100 µl | 5-7 µg |
Cat # | Name | Size | Price | |
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D3004-4-50 | DNA Elution Buffer | 50 ml | $32.00 | |
D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
D3004-1-100 | Genomic Lysis Buffer | 100 ml | $60.00 | |
D3004-2-100 | g-DNA Wash Buffer | 100 ml | $30.00 | |
D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
C2003 | Elution Plate | 2 Plates | $19.00 | |
C2002 | Collection Plate | 2 Plates | $22.00 | |
C2001 | Silicon-A Plate | 2 Plates | $129.00 |
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当然抗菌药中也不全都是抗生素,还有一部分是人工合成抗菌药,比如磺胺类(如SMZ)、喹诺酮类(如氧氟沙星)、硝基呋喃类(如痢特灵)、硝基咪唑类(如替硝唑)、恶唑烷酮类(如利奈唑胺)抗菌药。
欧美国家是否有相关权威的抗生素使用规范?如果有,请提供,最好是中英对照的,相关网站也可以
二、抗生素的种类:
由细菌、霉菌或其它微生物在生活过程中所产生的具有抗病原体不同的抗生素药物或其它活性的一类物质。自1943年以来,青霉素应用于临床,现抗生素的种类已达几千种。在临床上常用的亦有几百种。其主要是从微生物的培养液中提取的或者用合成、半合成方法制造。其分类有以下几种:
(一)β-内酰胺类:青霉素类和头孢菌素类的分子结构中含有β-内酰胺环。又有较大发展,如硫酶素类(thienamycins)、单内酰环类(monobactams),β-内酰酶抑制剂(β-lactamadeinhibitors)、甲氧青霉素类(methoxypeniciuins)等。
(二)氨基糖苷类:包括链霉素、庆大霉素、卡那霉素、妥布霉素、丁胺卡那霉素、新霉素、核糖霉素、小诺霉素、阿斯霉素等。
(三)酰胺醇类:包括氯霉素、甲砜霉素等。
(四)大环内酯类:临床常用的有红霉素、白霉素、无味红霉素、乙酰螺旋霉素、麦迪霉素、交沙霉素等、阿齐红霉素(阿奇霉素)。
(五)多肽类抗生素:万古霉素、去甲万古霉素、替考拉宁,后者在抗菌活性、药代特性及安全性方面均优于前两者。
(六):硝基咪唑类:甲硝唑、替硝唑、奥硝唑。
(七)作用于G-菌的其他抗生素,如多粘菌素、磷霉素、卷霉素、环丝氨酸、利福平等。
(八)作用于G+细菌的其他抗生素,如林可霉素、克林霉素、杆菌肽等.
(九)抗真菌抗生素:分为棘白菌素类、多烯类、嘧啶类、作用于真菌细胞膜上麦角甾醇的抗真菌药物、烯丙胺类、氮唑类。
(十)抗肿瘤抗生素:如丝裂霉素、放线菌素D、博莱霉素、阿霉素等。
(十一)抗结核菌类:利福平、异烟肼、吡嗪酰胺、利福布丁等。
(十二)具有免疫抑制作用的抗生素:环孢霉素等。
(十三)四环素类:包括四环素、土霉素、金霉素及强力霉素等。
在生活中,不少人认为抗生素是退烧药而随便使用,这是一种误区。其实不是所有的发热都是由细菌感染引起的,常见的伤风感冒通常由病毒所致,也发热,用抗生素毫无用处。而服用一般解热镇痛药大多可以奏效。患了病毒性感冒后,应到医院诊疗,只要加强护理,适当休息,多喝开水,给予易消化的饮食,通常会很快恢复健康。
俗话说,“是药三分毒”。滥用抗生素危害就更大了。首先,它会诱发细菌耐药。病原微生物为逃避药物,在不断地变异,耐药菌株也随之产生。目前,几乎没有一种抗菌药不存在耐药现象;其次,它会损害人体器官。抗菌药在杀菌的同时,也会造成人体损害,如影响肝、肾脏功能、胃肠道反应及引起再生障碍性贫血等;再次,它还会导致二重感染。在正常情况下,人体的口腔、呼吸道、肠道都有细菌寄生,寄殖菌群在互相拮抗下维持着平衡状态。如果长期使用抗菌药,敏感菌会被杀灭,而不敏感菌乘机繁殖,未被抑制的细菌、真菌及外来菌也可乘虚而入,诱发又一次感染。
是否使用抗生素治疗,首先应去医院做个血常规化验,查一下白细胞情况,搞清感冒的性质。如果是病毒性感冒,使用抗生素毫无用处,应使用抗病毒药物;如果感冒症状是由细菌感染造成的,则需使用抗生素。病人可根据感冒的症状进行自我初步判断:体温不高,痰是白色的,鼻水是清的,一般可初步判断为病毒性感冒;痰是黄色的,则可能是细菌感染。
小孩感冒应先到医院做血常规检查,确诊是病毒感冒还是细菌感染。如病毒感冒不需用抗生素治疗,如果是细菌感染,应在医生指导下选择适合儿童用的相对安全的抗生素治疗,如:青霉素类、头孢类抗生素。需要特别提醒的是,氨基糖苷类抗生素一定要慎用,因为它有耳毒性,有造成儿童耳聋的风险。
阅读抗生素的药物说明书时应重点关注适应证、用法、用量等细节。小儿用药剂量和大人不同,而有的家长不按说明服药,拿成人用的药量给孩子服用,以为只要对症就能治病。殊不知,有些抗生素对骨骼发育会产生抑制作用,成人可用,但孩子万万吃不得。需要特别提醒的是,儿童颗粒剂抗生素要用温开水调服,因为抗生素的稳定性不高,水温太热会影响药效。同时要避免用牛奶、饮料等服药。另外,患者在服用头孢类抗生素期间及治疗结束后72小时内(三天)应避免摄入含酒精的饮料,否则会引起戒酒硫样反应,人会有醉酒的感觉。还要避免吃奶酪,因为奶酪中有酪氨酸,会与药物发生反应。
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