Author Archives: Mike

  LB (Luria-Bertani) medium   Commercially available: or use the following recipe:   To 950 mls of ultrapure water, add   10g bacto-tryptone   5g bacto-yeast extract   10g NaCl.   15g bacto-agar, to solidify.   Mix, autoclave. PH to … Continue reading →

Stock solutions 70% Ethanol (1L) 736 ml 95% ethanol. Bring up to 1 L with nanopure water.Don’t autoclave.   EDTA 0.5 M   To prepare 500 ml, 0.5M EDTA pH 8.0: Add 84 g of disodium EDTA•2H2O to 300 ml … Continue reading →

Primers are delivered as dry pellets and can be storage at 4C. -Use sterile 1.5ml centrifuge tubes, add to each primer tube received 1ml nuclease free water using filter tips. -Adjust the concentration of your primers to a final of … Continue reading →

When working with RNA:   Nucleases (RNases) can destroy RNA and ARE EVERYWHERE, so you should:   Prepare an RNAse free working environment. Wear gloves while handling reagents and RNA samples to prevent RNase contamination from the surface of the … Continue reading →

| simple dilution  | serial dilution | VC=VC  method | molar solutions | percent  concentrations | molar – % conversions   1. Simple Dilution  (Dilution Factor Method)   A simple dilution is one in which a unit volume of a … Continue reading →

Basic PCR, 25 ul   Use tips and tubes nuclease free                                                                                     VOL                                         … Continue reading →

Reaction   EX: For a 20 ul total volume of reaction (15 ug DNA)                   5 ul DNA: Maximum DNA 1/3 of total volume          2 ul Buffer 10X: Maximum 1/10 of total volume          2 ul enzyme: NEB … Continue reading →

Digest good-quality genomic or plasmid DNA to completion with restriction enzymes. Use AT LEAST 1 ug  of genomic DNA per lane; 10 micrograms per lane is ideal.  If more than one DNA sample is to be compared, you should load … Continue reading →

Protocol used to concentrate poly(A)+ mRNA, RNA, DNA. Also to clean reactions from buffers and others that may interfere in dowstream reactions.   1. Add 1/10 volume 3 M NaAc (Sodium acetate) and 3 volumes of cool ethanol 95% (high … Continue reading →

  Vectors contain sequences that allow plasmid recovery (rescue) including genomic sequences from the transformed fungus. Sequences required for plasmid rescue are a bacterial origin of replication and an antibiotic resistance gene (e.g. Ampicillin or Kanamycin resistance). In addition, unique … Continue reading →