Glycogen (Oyster)

Glycogen is a highly purified polysaccharide from oysters and can be used as a carrier for nucleic acid precipitation thereby replacing tRNA, linear polyacrylamide or sonicated DNA. Glycogen is an inert material compatible with most molecular biology procedures: PCR*, DNA sequencing, restriction digestion, ligation, competent cell transformation, cDNA synthesis, DNA labeling by kinase reactions and random priming, in vitro transcription/translation, gel electrophoresis, etc. Quality Control: Nucleic Acids Precipitation Assay: >95% of total radioactivity of 5pg [32P] calf thymus DNA was found in the precipitate after centrifugation. 5pg of [32P]-labeled calf thymus DNA were dissolved in 500ul TE buffer with 0.4M LiCl. 1ul glycogen solution (20ug) was added and then precipitated with 1.2ml of 96% ethanol at -20 degrees C, stored for 3 hours at -20 degrees C and centrifuged. Protease Assay: No detectable degradation of 0.6% FTC-casein was observed after incubation with 1000ug of glycogen in 200ul of reaction buffer for 16 hours at 37 degrees C. Ribonuclease Assay: No detectable radioactivity was released into the trichloroacetic acid-soluble fraction after incubation of 200ug of glycogen with 1ug of E. coli [3H]-RNA in 50ul of reaction buffer. Nucleic Acids Assay: After precipitation of the T4 polynucleotide kinase reaction mixture containing 200ug glycogen and washing with ethanol, radioactivity in the precipitate did not exceed that in the negative control. 200ug glycogen were incubated with 6pmol of [gamma-32P]-or [gamma-33P]-ATP and 10 units of T4 polynucleotide kinase for 20 minutes in 40ul of reaction buffer. Nicking Activity Assay: No detectable conversion of supercoiled DNA to nicked DNA was observed after incubation of 200ug of glycogen with 1ug of pUC19 DNA in 50ul of reaction buffer for 4 hours at 37 degrees C. Labeled Oligonucleotide (LO) Assay: No detectable degradation of oligonucleotides was observed. Single-stranded and double-stranded [32P]-labeled oligonucleotide were incubated with 50ug of glycogen for 16 hours in 5 restriction buffers at 37 degrees C and 55 degrees C. *The Polymerase Chain Reaction (PCR) process is covered by U.S.patents owned by Hoffman-La Roche. Storage and Stability: May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.