DNA实验技术

Preparation of Silica1. Suspend 5 g of silica (Sigma S-5631) in 50 ml of PBS.2. Allow the silica to settle for 2h.3. Discard the supernatant containing fine particulate matter.4. Repeat steps 2 and 3. ...

competent cells (DH5alpha at competency of 108 cfu/µl of pUC18). The transformation procedure it self is very similar to a standard CaCl2 transformation. Place 100 µl of TB buffer in a tube on ice. Me ...

Recovery of DNA from LMP (Low Melting Point) Agarose Gel1. Separate DNA fragments through an LMP agarose gel containing ethidium bromide (0.5 microgram/ml). 2. Detect DNA by irradiating the gel with l ...

Preparation of linear polyacrylamide used as carrier in ethanol precipitation of nucleic acidsLinear polyacrylamide can be used as an efficient neutral carrier for precipitating nucleic acids with eth ...

I tried to clean a restriction product 10kb before ligation by a gel extraction method (Qiagen gel extraction kit). But the concentration was too low to do a ligation. I used to clean restriction prod ...

Rapid elution of DNA from agarose gels James Movius Hahn LabLast modified Sun Nov 1 1998A method of quickly purifying agarose gel DNA fragments for use in subsequent reactions such as further restrict ...

OverviewProcedure 1. Preheat the CTAB Isolation Buffer at 60℃.2. Grind 2 g of fresh leaf tissue to a powder in Liquid Nitrogen in a chilled mortar and pestle.3. Scrape the powder into a chilled 50 ml ...

Preparation of plasmid DNA : a modified mini alkaline-lysis/PEG precipitation procedure. ABI; 1995MaterialsGTE buffer (50mM glucose 25mM Tris-HCl (pH8.0) 10mM EDTA (pH 8.0)) (200µl/tube) 0.2N NaOH / 1 ...

CTAB TECHNIQUE / Method / Schedule / Protocol (JPB) FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES(see also if both DNA and RNA are needed)Reagents neededCTAB buffer2% CTAB 20gm C ...

Hidoes anyone know what is the best temperature and time for Isoprop precipitation of DNA? I usually do at room temperature and centrifuge immediately. Can I improve this? And would it ma ...

1. Pour a vertical acrylamide gel using TEA buffer. A 4 % non denaturing gel is correct for most applications.2. Run out DNA fragments. For fragments greater than 500 bp run the xylene cyanol dye to t ...

Phenol-based Method for the Isolation of DNA Fragments from Low-Melting Temperature AgaroseReference:Favre D. 1992. Biotechniques vol. 13 1.Cut out slice containing DNA smallest size possible. 2.Estim ...

NOTE: THIS IS FINE FOR SOUTHERNS BUT NOT FOR SCREENING BY PCR. (from Ruixia 7/99 from protocol by Stef Oehen 7/94). 1. Put 1 cm tail in 1.5 ml microcentrifuge tube.2. Add 500µl Tail Buffer and 30 µl P ...

1) Digest DNA with two restriction enzymes such that one end is ExoIII susceptible (blunt or 5' overhang) and the other is ExoIII resistant (5' recessed). The ExoIII susceptible end should be such tha ...

CAT ASSAY 1. Transfer cells to a 15 ml tube. 2. Add 5 ml TBS- to flasks shake & pour into tubes. 3. Spin down the cells @ 1k rpm for 5'. 4. Resuspend in 1 ml TBS- . 5. Transfer to 1.5 ml tubes. 6. Spi ...

The following guidelines should be taken into account when designing modified oligonucleotides. 1.Sequence Length - SYNTHEGEN can synthesize oligonucleotides from 5 to 110 bases in length. Most sequen ...

Used to removed unincorporated nucleotides from labelling reactions.Prepare Sephadex G-50 (medium) by adding appropriate amount of dry beads to 100 ml TE buffer such that the beads will swell to 50 ml ...

1. Pick single colony and inoculate 5 ml of LB broth containing 200 g/l ampicillin or 1mg/5ml. Optional: Use a 15ml conical tube with a loosened cap and a piece of tape to hold it in place. Shake at 2 ...

Author: Long-Cheng Li Source: Protocol Online Abstract: Simplified method for preparing G A ladder run along with footprinting reaction. It's much simple than the original Maxima-Gilbert sequencing re ...

Protocol for extracting DNA from ES Cells starting from the 96-well plate but processing in an eppendorf tube to recover more of the DNA . NOTE- THIS TAKES A LOT OF TIME if you do the whole plate this ...