Friday, November 21, 2003 Description The original maize DNA miniprep protocol is used extensively for many plant species and different tissues. This slightly modified version is acceptable for most DNA extractions. The procedure has the a...
Description Procedure 1. Place 0.5 to 2 g (1 g is ideal) of plant tissue in a mortar and add an excess of liquid nitrogen. As the liquid nitrogen evaporates, grind the tissue thoroughly into a fine powder. 2. Transfer the plant powder into...
Description This protocol describes a method for isolating DNA from plant tissue. Procedure 1. Preheat the CTAB Isolation Buffer at 60°C. 2. Grind 2 g of fresh, leaf tissue to a powder in Liquid Nitrogen in a chilled mortar and pestle. 3....
Friday, November 21, 2003 Description Cyanine Dye Purification Protocol Procedure Preparation of salt buffers The buffers are used for both HPLC purification and Sep-Pak purification. High salt buffer (HSB): 1M triethylammonium acetate, pH...
Friday, November 21, 2003 Description Preparation of Fluorescent DNA Probe from HUMAN mRNA or Total RNA using Direct Incorporation Procedure I. Preparing fluoresenctly labeled cDNA (probe): To anneal primer, mix 2ug of mRNA or 50-100mg tot...
Friday, November 21, 2003 Description Hybridization of Arrays Procedure Directions for fluorescent DNA probes: 1. Final probe volume should be 12-15uL, at 4X SSC, containing competitor DNA etc. as required (e.g., polydA, CoT1 DNA for a hum...
Friday, November 21, 2003 Description Post-Processing of Arrays Procedure Mark boundaries of array on back of slide using diamond scriber. Array will become invisible after post-processing. Fill bottom of humid chamber with 100 ml 1X SSC....
Friday, November 21, 2003 Description Protocol for Reverse Transcription and Amino-allyl Coupling of RNA Procedure A. RT Reaction 1. To anneal primer, mix 1-2 mg mRNA with 5 ug of anchored oligo-dT [(dT)20-VN] (Operon, HPLC purified) in a...
Friday, November 21, 2003 Description Resuspend the contents of one pack of the monofunctional NHS-ester Cy3 or Cy5 dye (mono-functional Cy3 or Cy5 reactive pack, Amersham catalog PA 23001 and PA 25001) in 72 ul DMSO. Aliquot 16 x 4.5 ul a...
Friday, November 21, 2003 Description Poly-l-Lysine coating of slides Procedure 1. Place slides in slide racks. Place racks in chambers. 2. Prepare CLEANING SOLUTION: Dissolve 70 g NaOH in 280 mL ddH2O. Add 420 mL 95% ethanol. Total volume...