Highlights
- Quick & Easy: Simple 20 minute procedure.
- Highest Yield: Recover 3x more DNA.
- Ultra-Pure: Ready for qPCR, Next-Gen Sequencing, arrays, etc.
Description
| Elution Volume | ≥ 35 µl |
|---|---|
| Equipment | Water bath or heat block (55°C), microcentrifuge, and vortex. |
| Purity | High quality DNA is ready for all sensitive downstream applications such as PCR, endonuclease digestion, Southern blotting, genotyping, Next-Gen sequencing, bisulfite conversion, etc. (A260/A230≥ 2.0). |
| Size Range | Capable of recovering genomic and mitochondrial DNA sized fragments > 50 kb. If present, parasitic, microbial, and viral DNA will also be recovered. |
| Workflow | Utilize a Proteinase K Digestion and Zymo-Spin Column for effective recovery of DNA. |
| Yield | The DNA binding capacity of the column is 25 µg. Typically, mammalian tissues yield: 1-3 µg DNA per mg skeletal, heart, lung, and brain tissues and 3-5 µg DNA per mg liver and kidney. Human whole blood will yield 3-7 µg DNA per 100 µl blood sampled. |
Q1: What is the difference between the digestion buffers? Can one buffer be used for all sample types?
The BioFluid & Cell Buffer (Red) is designed to allow for rapid Proteinase K digestion with easy-to-lyse samples (e.g. mammalian cells and biological fluids).The Solid Tissue buffer (Blue) can be used for any sample type and requires a longer PK digestion time compared to using the BioFluid & Cell Buffer (Red).
Q2: 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 recovers more DNA with higher purity compared to the Quick-DNA Kits.
Q3: Can Proteinase K digestion be performed overnight in DNA/RNA Shield?
Yes, samples can be digested overnight. Make sure to follow the appendix (page 8 appendix B) for processing liquids samples in DNA/RNA Shield and incubate at room temperature.
Q4: 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 aliquoting for purification.
Q5: Can the Quick-DNA Plus kit be used with bacterial samples?
E.coli cells are easy-to-lyse and can be processed directly with the Biological Fluids & Cells protocol. For an all-inclusive kit with any type of microbes (including tough-to-lyse), use any of Zymo Research’s Environmental Kits (e.g. Quick-DNA Fungal/Bacterial, Quick-DNA Fecal/Soil, ZymoBIOMICS DNA, etc.).
Q6: Can I use Quick-DNA Plus to clean-up previously isolated DNA?
No, the kit is designed for direct use with biological samples. For clean-up of previously isolated DNA, please use the Genomic DNA Clean & Concentrator or the DNA Clean & Concentrator kits.
Q7: 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.
Q8: 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.
This kit is easy to use and provides a high quality yield.
-M.B. (Syracuse University)
“I was very impressed with the purity of the DNA.”
-C.V. (UCLA)
“Improved yield than Bioline and Qiagen Kits.”
-F.C. (Manchester Metropolitan University)
Read More| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| C1001-50 | Collection Tubes | 50 Pack | $15.00 | |
| D3004-2-50 | g-DNA Wash Buffer | 50 ml | $18.00 | |
| D3004-4-1 | DNA Elution Buffer | 1 ml | $11.00 | |
| D3004-4-10 | DNA Elution Buffer | 10 ml | $14.00 | |
| D3004-2-200 | g-DNA Wash Buffer | 200 ml | $54.00 | |
| D3004-4-50 | DNA Elution Buffer | 50 ml | $32.00 | |
| D3004-5-50 | DNA Pre-Wash Buffer | 50 ml | $26.00 | |
| D3004-5-30 | DNA Pre-Wash Buffer | 30 ml | $21.00 | |
| D3001-2-20 | Proteinase K w/ Storage Buffer Set | 20 mg | $44.00 | |
| D3001-2-5 | Proteinase K w/ Storage Buffer Set | 5 mg | $21.00 | |
| D4068-1-12 | BioFluid & Cell Buffer (Red) | 12 ml | $24.00 | |
| D4068-1-45 | BioFluid & Cell Buffer (Red) | 45 ml | $59.00 | |
| D4068-2-22 | Solid Tissue Buffer (Blue) | 22 ml | $38.00 | |
| D4068-2-6 | Solid Tissue Buffer (Blue) | 6 ml | $12.00 | |
| D4068-3-25 | Genomic Binding Buffer | 25 ml | $28.00 | |
| D4068-3-85 | Genomic Binding Buffer | 85 ml | $95.00 | |
| C1103-50 | Zymo-Spin IC-XM Columns | 50 Pack | $63.00 | |
| C1104-50 | Zymo-Spin IIC-XLR Columns | 50 Pack | $66.00 |
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TBST中含有Tris-Hcl，NaCl，Tween20这三种物质，是做WESTERNBLOT中常用的一种缓冲液。
TBST缓冲液的配制
1000ml×TBST的配置
先称量NaCl40g，倒入烧杯中，加DDW蒸馏水400ml,再称量NaCl47.6g，倒入刚才的那个烧杯中（PS：由于NaCl的量太多，一次称量不方便，所以分两次称量，且易于溶解）。往烧杯中加入Tris—HCl缓冲液100ml，最后加（吐温20）5ml,转入1000ml容量瓶中，在定容，转移即可。
TBST缓冲液的应用：
1.主要用于免疫组化和原位杂交，酶联免疫等实验中，清洗免疫印。
2.迹膜；
注意事项：
1.TBST缓冲液，PH7.2-7.5；
2.颜色为无色透明液体；
3.为了您的安全和健康，请穿实验服并戴防护手套操作；
1 mmol/LEDTA(pH 8.0)
因为含有以上两种物质，所以称为TE。
配制分三步：
1)1 M Tris-HCl (pH 8.0) 50 ml的配制：称取Tris碱6.06 g，加超纯水40 ml溶解，滴加浓HCl约2.1 ml调pH至8.0，定容至50 ml。
2)0.5 M EDTA（pH 8.0）50 ml的配制：称取EDTA-Na2·2H2O 9.306 g，加超纯水35 ml，剧烈搅拌，用约1 g NaOH颗粒调pH至8.0，定容至50 ml。（EDTA二钠盐需加入NaOH将pH调至接近8.0时，才会溶解。）
3)1×TE（10 mM Tris-HCl，pH 8.0；1 mM EDTA，pH 8.0）的配制：
作用：
TE缓冲液是弱碱性,对DNA的碱基有保护性,(DNA在它是的稳定性较好,不易破坏其完整性或产生开环及断裂),包括提取好的DNA也要放在TE缓冲液是保存. 10mMTris-Hcl，pH有7.47.68.0三种。
EDTA调到8.0是为了更好溶解，其他只要调到相应pH就可以。Tris在7－8附近缓冲能力很强，所以加8.0的EDTA下去后，不会改变pH。
在缓冲溶液中加入少量强酸或强碱，其溶液pH值变化不大，但若加入酸，碱的量多时，缓冲溶液就失去了它的缓冲作用。这说明它的缓冲能力是有一定限度的。
缓冲溶液的缓冲能力与组成缓冲溶液的组分浓度有关。0.1mol·L-1HAc和0.1mol· L-1NaAc组成的缓冲溶液，比0.01mol·L-1HAc和0.01mol·L-1NaAc的缓冲溶液缓冲能力大。关于这一点通过计算便可证实。但缓冲溶液组分的浓度不能太大，否则，不能忽视离子间的作用。
组成缓冲溶液的两组分的比值不为1∶1时，缓冲作用减小，缓冲能力降低，当c（盐）/c（酸）为1∶1时△pH最小，缓冲能力大。不论对于酸或碱都有较大的缓冲作用。缓冲溶液的pH值可用下式计算：
此时缓冲能力大。缓冲组分的比值离1∶1愈远，缓冲能力愈小，甚至不能起缓冲作用。对于任何缓冲体系，存在有效缓冲范围，这个范围大致在pKaφ（或pKbφ）两侧各一个pH单位之内。
弱酸及其盐（弱酸及其共轭碱）体系pH=pKaφ±1
弱碱及其盐（弱碱及其共轭酸）体系pOH=pKbφ±1
例如HAc的pKaφ为4.76，所以用HAc和NaAc适宜于配制pH为3.76～5.76的缓冲溶液，在这个范围内有较大的缓冲作用。配制pH=4.76的缓冲溶液时缓冲能力最大，此时（c（HAc）/c（NaAc）=1。
制备
为了配制一定pH的缓冲溶液，首先选定一个弱酸，它的pKaφ尽可能接近所需配制的缓冲溶液的pH值，然后计算酸与碱的浓度比，根据此浓度比便可配制所需缓冲溶液。
以上主要以弱酸及其盐组成的缓冲溶液为例说明它的作用原理、pH计算和配制方法。对于弱碱及其盐组成的缓冲溶液可采用相同的方法。
缓冲溶液在物质分离和成分分析等方面应用广泛，如鉴定Mg2+ 离子时，可用下面的反应：
白色磷酸铵镁沉淀溶于酸，故反应需在碱性溶液中进行，但碱性太强，可能生成白色Mg（OH）2沉淀，所以反应的pH值需控制在一定范围内，因此利用NH3·H2O和NH4Cl组成的缓冲溶液，保持溶液的pH值条件下，进行上述反应。
常用缓冲液配制
枸橼酸-磷酸氢二钠
甲液:取枸橼酸21g或无水枸橼酸19.2g,加水使溶解成1000ml，置冰箱内保存。
乙液：取磷酸氢二钠71.63g，加水使溶解成1000ml。
取上述甲液61.45ml与乙液38.55ml，混合，摇匀，即得。
氨－氯化铵缓冲液
取氯化铵1.07g，加水使溶解成100ml， 再加稀氨溶液(1→30)调节pH值至8.0，即得。
氨－氯化铵缓冲液
取氯化铵5.4g，加水20ml溶解后，加浓氨溶液35ml，再加水稀释至100ml，即得。
醋酸－醋酸钠缓冲液
取无水醋酸钠20g，加水300ml溶解后,加溴酚蓝指示液1ml及冰醋酸60～80ml，至溶液从蓝色转变为纯绿色，再加水稀释至1000ml，即得。
醋酸－醋酸钠缓冲液
取醋酸钠18g,加冰醋酸9.8ml,再加水稀释至1000ml，即得。
醋酸－醋酸钠缓冲液
取醋酸钠54.6g，加1mol/L醋酸溶液20ml溶解后，加水稀释至500ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵7.7g，加水50ml溶解后，加冰醋酸6ml与适量的水使成100ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵77g，加水约200ml使溶解，加冰醋酸57ml,再加水至1000ml，即得。
醋酸－醋酸铵缓冲液
取醋酸铵100g，加水300ml使溶解，加冰醋酸7ml,摇匀，即得。
磷酸盐缓冲液
取0.2mol/L磷酸二氢钾溶液250ml,加0.2mol/L氢氧化钠溶液118ml，用水稀释至1000ml，即得。
当往某些溶液中加入一定量的酸和碱时，有阻碍溶液pH变化的作用，称为缓冲作用，这样的溶液叫做缓冲溶液。弱酸及其盐的混合溶液（如HOAc与NaOAc），弱碱及其盐的混合溶液（如NH3·H2O与NH4Cl）等都是缓冲溶液。
参考资料：http://baike.baidu.com/view/901429.htm
求采纳为满意回答。
