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FastDigest PstI
描述
| 5' | C | T | G | C | A ↓ | G | 3' | ||||
| 3' | G ↑ | A | C | G | T | C | 5' |
Thermo Scientific FastDigest PstI restriction enzyme recognizes CTGCA^G site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: BspMAI.
Thermo Scientific FastDigest PstI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. See Reaction Conditions for FastDigest Enzymes for a table of enzyme activity, heat inactivation and incubation times for this and other FastDigest restriction enzymes. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.
For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.
Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.
Features
• 100% activity of all FastDigest enzymes in the universal buffer
• 100% buffer compatibility with downstream applications
• Complete digestion in 5–15 minutes
• Direct loading on gels
规格
| Compatible Buffer: | 10x FastDigest Buffer/FastDigest Green Buffer |
|---|---|
| Enzyme: | Pst I |
| Methylation Sensitivity: | Not CpG methylation-sensitive, Not dam methylation-sensitive, Not dcm methylation-sensitive |
| Optimal Reaction Temperature: | 37° C |
| Product Line: | FastDigest |
| Product Size: | 800 reactions |
| Sensitive to Heat Inactivation: | No |
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文献和实验伤寒杆菌组氨酸转运操纵子为起点进行连续步行。以M13mpl8RF DNA 为载体。用PstI和AraI酶切基因组DNA ,PstI和XmaI酶切载体DNA ,然后连接基因组片段和载体片段,用根据基因组DNA 序列设计的特异引物和载体的通用引物进行扩增,由于非特异片段没有单特异引物结合的位点,即使有载体连到非特异片段,也无法得到大量扩增,而使特异片段得到有效扩增。 1.4.2 利用接头的PCR 王新国等利用衔接头的方法,设计了位于单链DNA 两端互补的颠倒末端重复序列,增加了反应的特异
Ligation of DNA fragments by homopolymer tailing
with the restriction endonuclease PstI (recognition site: CTGCAG) which will produce the 3’- extension TGCA at both termini. If the PstI digested vector DNA is then homopolymertailed with dGTP, and consequently the target DNA fragments are tailed with the complementary
Subcloning: restriction enzyme digests, ligation
diphosphates and triphosphates and selectivity identical to that of the wild-type receptor. Biochemical Pharmacology 59, 791-800). You will isolate a 1.4kb PstI-BamHI DNA fragment (insert) containing the amplified P2X2 sequence (that you generated using
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