Highlights
- Simple: Quickly and easily rescue plasmid from yeast.
- Efficient Isolation: Works well with low-copy and hard to isolate plasmids.
- High-Quality: Isolated plasmid DNA is ideal for molecular biology techniques, such as PCR, transformation, hybridization, etc.
Description
| Applicable For | Plasmid DNA is well suited for downstream applications such as PCR, transformation, hybridization and other sensitive applications |
|---|---|
| Elution Volume | ≥ 35 µl |
| Equipment | Microcentrifuge & Heat Block/Bath |
| Processing Volume | ≤ 1.5 ml of Culture |
| Sample Source | Cell Culture (Colonies/Patches or Liquid Culture) |
| Size Range | No Size Limit |
| Yield | Typically between 0.01-0.3 ng for most 2 µ based plasmids from 1.5 ml overnight cultures |
Q1: What is the difference between Zymoprep Yeast Plasmid Miniprep I and Miniprep II?
Both the Zymoprep Yeast Plasmid Miniprep I and II utilize the same chemistry for lysis; however, Miniprep I uses isopropanol precipitation and Miniprep II utilizes a column for purification. The Miniprep II allows for consistent yield and purity; and samples can be concentrated to a low elution volume.
Q2: What is the typical plasmid yield?
Typically, between 0.01 - 0.3 ng for most 2 µ based plasmid from 1.5 ml overnight cultures.In order to generate more plasmid, the plasmid is typically transformed into E. Coli , cultured, and isolated using a traditional E. Coli plasmid prep.
Q3: Can this kit be used to isolate linear plasmid DNA?
Yes
Q4: If I’m using stationary phase yeast cells, what can I do to improve sample lysis?
We generally recommend working with fresh or early log phase cells, which are easier to lyse. For stationary phase cells, user optimization is necessary, and we recommend increasing digestion to > 1 hour and/ or increasing the amount of Zymolyase.
Q5: What strains are these kits compatible with?
Any strains susceptible to Zymolyase, which includes the following fungal genera: Asbya, Kloekera, Candida, Kluyveromyces, Debaryomyces, Lipomyces, Eremothecium, Metschikowia,Endomyces, Pichia, Hansenula, Pullularia, Hanseniaspora, Saccharomyces, Saccaromycodes, Saccharomycopsis, Schizosaccahromyces, Torulopsis
| Cat # | Name | Size | Price | |
|---|---|---|---|---|
| D2001-1-15 | Solution 1 Digestion Buffer | 15 ml | $18.00 | |
| D2001-2-15 | Solution 2 Lysis Buffer | 15 ml | $18.00 | |
| D2001-3-15 | Solution 3 Neutralizing Buffer | 15 ml | $18.00 |
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② 应取pH8.0,这样可使核苷酸带较多负电荷,利于吸附于阴离子交换树脂柱。虽然pH 11.4时核苷酸带有更多的负电荷,但pH过高对分离不利。
③ 当不考虑树脂的非极性吸附时,根据核苷酸负电荷的多少来决定洗脱速度,则洗脱顺序为CMP>AMP> GMP > UMP,但实际上核苷酸和聚苯乙烯阴离子交换树脂之间存在着非极性吸附,嘌呤碱基的非极性吸附是嘧啶碱基的3倍。静电吸附与非极性吸附共同作用的结果使洗脱顺序为:CMP> AMP > UMP >GMP。
1. Buffer中离子浓度过大,甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高,也是一个原因
我使用的是1%琼脂糖凝胶,电泳缓冲液为0.5×TBE。
请问各位大虾,在不影响实验结果的前提下,这样的含EB的电泳缓冲液可以反复使用多少次?
谢谢^_^
500ml
2。阴极缓冲液 ( 10 × ) :将 60.55g Tris ,89.58g Tricine 及 5g SDS 溶于400ml 蒸馏水中,加水至终体积为500ml
使用时稀释至1×的缓冲液,电泳槽内槽加入阴极电泳缓冲液,外槽加入阳极电泳缓冲液。形成Trcine-SDS-PAGE电泳系统。
如果你是制胶或者做电泳缓冲液的话,不用灭菌。
Tris-Glycine/SDS;
MOPS/SDS;
Bis-Glycine/SDS等不同的缓冲液、预混液等。
不知道这几种缓冲液有什么异同呢?
比如如果是Invitrogen的预制胶-预混液电泳系统,每种预制胶有适配缓冲液,但是如果用其他的缓冲液似乎也可以得到不错的结果,那么是否说明电泳缓冲液只要满足了缓冲ph范围,其他的不是特别重要呢?
此外,附加提问是,PVDF膜在转膜的时候,转膜缓冲液中不需要加入甲醇,那么是加入甲醇转膜效果好还是不加好呢?
谢谢!

