DNA提取与纯化

1. Separate DNA fragments in an agarose gel cast with 0.5 mg/ml Ethidium bromide. Locate bands with a hand-held long-wave UV lamp.2. Slice the gel with a razor blade above and below the bands of inter ...

Equipment and reagents1. Phenol2. TE bufferpH 8.0 (10 mM Tris-HClpH 8.0;1 mM EDTApH 8.0)3. 24:1 (v/v) chloroform-isoamyl alcohol4. 3 M potassium acetate pH 5.5prepared by adding glacial acetic acid t ...

Author: Suzanne GerttulaDate: March 141994 Digest between 1 and 10 ug Genomic DNA. I digest at 37 C for about six hours over the course of a day. Spin down briefly heat to 65 C for 10 min and then chi ...

1. Spin down 1.5 ml of overnight culture in 2ml or 1.5ml tube for 1 minute on high. 2. Asprirate supernatant and resuspend cell pellet in 100 µl Solution I (25 mM Tris-HCl pH 8.0 10 mM EDTA). 3. Add 2 ...

PURPOSE: TolocateaparticularsequenceofDNAwithinacomplexmixture(locateonegenewithinanentiregenome) SeparatemixtureofDNAsequencesonagelprobewithspecificDNAsequence(gene) Otherrelatedblottingtechniques: ...

1. Add an equal volume (equal to sample volume) of P/C to sample.2. Mix (shake don't vortex).3. Take aqueous (upper) layer. (If dirty sample repeat Ph/Chl step until interface is fairly clean).4. Add ...

IntroductionMethods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts available on the internet. A commonly occurring theme on t ...

This is a scaled up version of the Baron protocol which has been modified to achieve purity comparable to CsCl preps. I have not sorted out strain variations yet but HB101 works well.1、SolutionsSoluti ...

There are many methods for DNA preparation without using kits.Below are a few minipreps.These protocols may be scale up to midi or maxi prep if necessary.1.Ammonium Acetate Method 1)Spin down cell and ...

1.Prepare or obtain Buffered phenolpH 8.0.Add 2 crystals of 8-Hydroxyquinoline to prevent oxidation.This also identifies the organic phase as yellow-colored.2.Combine DNA sample with an equal volume o ...

1、Grow an overnight culture from a single colony in 2 mls LB antibiotic.2、Transfer 1.5 mls of culture to eppendorf tube.Spin for 1 minute on high and remove supernatant.3、Add 700 µl STET. Add 25 µl ...

1、Put a 0.5-1.5 CM MOUSE TAIL SNIP in a 1.5 ml eppindorf tube and store frozen until digestion.Make certain that the eppindorf tubes seal wellotherwise your samples may leak out during the shaking ste ...

Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brush ...

1.grow up single colony in 1.5 ml LB/antibiotic ovn @37℃ .2.pour into tube spin for 30 sec .3.decant supernatant and resuspend pellet in 100 ml lysis solution .4.add 200 ml alkaline SDS vortex well .5 ...

This is a standard large scale prep.for plasmid DNA which gives a yield of 0.5-1.0mg.I have made some minor changes to the MHB protocol.Solutions Solution IIIIII from protocol D.1.Tris/EDTA pH 7.5 (op ...

A. Culture Growth: 1.For BAC isolation for shotgun library construction: Pick a smear of colonies and inoculate 3 ml TB medium containing the appropriate antibiotic. Grow the culture for 8 hours 37 de ...

1、The BAC clone from glycerol culture sublibrary is innoculated into 3 ml of LB with 20µg/ml chloramphenicol and cultured at 37°C with shaking for 16hrs. The liquid bacterial culture is transferred in ...

点击浏览该文件

After an aliquot of the PCR mixture is analyzed on an agarose gel the remainder of the reaction is concentrated by ethanol precipitation resuspended in buffer and subjected to a simultaneous fill-in/k ...

1、A smear (rather than a single colony) of BAC colonies are picked and transferred into a 12X75 mm Falcon tube containing 3 ml of LB medium (10 g Bacto-Tryptone 5 g Bacto-yeast extract and 10 g NaCl i ...