DNA基础知识

CTAB TECHNIQUE / Method / Schedule / Protocol FOR DNA ISOLATION / DNA EXTRACTION FROM PLANT LEAF / LEAVES SAMPLES (see also DNA RNA double isolation procedure if both DNA and RNA are needed) Reagents ...

Use from 0.01 - 0.1 gram plant material. Grind the plant material with liq. N2 in a mortar. We normally use some alumina to crush hard tissue. Transfer the ground tissue to a eppendorf tube. Add 1 ml ...

Plant_Leaf_DNA_Extraction.pdf

Plant_Leaf_DNA_Extraction.pdf

Preheat Extraction Buffer at 60°C. Weigh 100 mg of fresh leaf tissue and grind it to powder in Liquid Nitrogen in a chilled mortar and pestle. Add 0.5 ml of TE buffer to the tube. Centrifuge at 500 g ...

Grow Rhizobium cells in 5 ml of YEMB at 200 rpm till the O.D (600 nm) reaches 0.6-0.8. Pellet the cells by centrifugation at 10000 rpm for 15 mins. Wash the pellet with TE buffer (10T/1E). Dissolve th ...

Phenol Chloroform Absolute ethanol 70% ethanol TE buffer (10T/1E) pH 8.0 10% SDS 50 mg/ml lysozyme 5 M NaCl ProcedureMacerate Cyanobacteria in TE Buffer (10T/1E) Pellet the cells by centrifugation for ...

Procedure:1) Use Pharmacia #27-4564-01. With clean flamed scissors and forceps weigh 0.25 g/50 ml conical and add 50 ml/conical of 0.02 M Tris pH 7.6. Allow to dissolve over several days at 4°C.2) ...

PCR-based methods are widely used in plants and animals for marker-assisted breeding and high-resolution mapping. These studies require analysis of large number of samples thus a DNA extraction method ...

Take an oral smear Dissolve in 50 ml H2O through vigorous shaking Pellet cells 5 min at 4000 rpm Re-suspended the pellet in 400 μl Lysis buffer (50 mM TRIS-Cl pH 8 10 mM EDTA 2% SDS) Incubate for 5 mi ...

A nested series of deletions can be produced by: cutting plasmid DNA (2.5ug per time point see below) with an enzyme leaving a blunt (eg EcoRV) or 5'' overhand (eg EcoRI) on the side that the deletion ...

Procedure:1) Need to cut 10 µg of plasmid in two spots (‘A’ and ‘B’) in polylinker. ‘A’ cut is with a restriction enzyme which gives a 3’ recessed end (exonuclease sensitive) next to insert. Since ...

0.25M HCl (1L)20.66mL HCl 979.34mL dH2O *add acid to wateundefined 1M ammonium acetate 0.02M NaOH (1L)77.08g ammonium acetate 0.8g NaOH Fill with dH2O 0.5X SSC 0.1% SDS (1L)25mL 20X SSC 10mL 10% SDS 965mL dH ...

Smith and Summers 1980. Anal. Biochem. 109:123-129.Digest 10-15 µg genomic DNA with desired restriction enzyme overnight at 37oC. Run digest on a 1% TBE agarose gel at 100V until 1st blue dye reaches ...

Genomic DNA Digest10 µl DNA (10 µg) 3 µl 10X Buffer 1-2 µl Restriction Enzyme (40 units) 3 µl 10mM Spermidine-HCl 3 µl 1 mg/ml BSA 10 µl H2O Total volume: 30 µlNote: Use DNA from a standard tail prep ...

SOLUTIONSLysis Buffer 0.1 M Tris pH 8.0 0.2 M NaCl 5 mM EDTA

Principle: Refer to Southern Transfer with Zetabind membrane. The NaOH transfer may be a more efficient transfer method for larger sized fragments. It is the method of choice for pulsed-field gels con ...

I. Preparing worm genomic DNA: requires 1-2 days to seed agarose plates a few days for the worms to grow 1-2 days to prep the DNA1. Seed large agarose plates with HB101. Agarose is preferred over agar ...

MaterialsHot Probe dNTP Mix:25 mM dATP25 mM dTTP25 mM dGTP2.5 mM dCTPHybridization Solution:5X SSC0.5% (w/v) Blocking Reagent0.1% (w/v) N-lauroylsarcosine Na-salt0.02% (w/v) SDS50% FormamideBlot Wash ...

IRS-1 Probe DNA Prep 1. From glycerol stocks (in �80ºC freezer) grow up bacteria. Use amp LB. 2. Use a Qiagen prep to purify the DNA. 3. Perform the following digest: ...