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LabBench Today                                                         September No.3

   

   

Procipitate Applications

Human Fibroblasts

Stephanie M Cohen, Terrence S Furey, Norman A Doggett, and David G Kaufman. Genome-wide sequence and functional analysis of early replicating DNA in normal human fibroblasts BMC Genomics.2006;7: 301

Procipitate™ was used in a study that replicated mammalian genomic DNA during the S phase. Scientists created a cosmid library containing DNA enriched in sequences to replicate early in the S phase of normal human fibroblasts. Next the sequencing and alignment of the clone ends to the human genome occurred. Cosmid DNA template was purified in 96-well format using ProPrep™ and ProCipitate™ from Biotech Support Group.

Bacterial artificial chromosome (BAC) vector

J M Kelley, C E Field, M B Craven, D Bocskai, U J Kim, S D Rounsley, and M D Adams. High throughput direct end sequencing of BAC clones.Nucleic Acids Res. 1999 March 15; 27(6): 1539-1546.

Bacterial artificial chromosome (BAC) vector generated libraries are popular for forming clone sets in high throughput genomic sequencing projects primarily because of their high stability. A low cost, efficient, 96 well procedure for BAC end sequencing for creating BAC end sequences from human and Arabidoposis libraries with high accuracy is described in this article. To develop a basic alkaline lysis filter plate that is processed in a 96 well template purification procedure yielding BAC DNA for several sequencing reactions and a DNA fingerprint, researchers used Procipitate™ from Biotech Support Group. Procipitate™ bound to proteins, removed it and helps with proper flow through the filter plates.

 

 

Procipitate™ References 
 

Roberto A. Rodríguez, Lauren Thie, Christopher D. Gibbons, Mark D. Sobsey. Reducing the effects of environmental inhibition in quantitative real-time PCR detection of adenovirus and norovirus in recreational seawaters. Journal of Virological Methods;2012:181(S1):43-50

Zhang, L., Lv, X., Tong, X., Jia, H. and Li, Z.Study on molecular mechanism for improving neural regeneration after repair of sciatic nerve defect in rat by acellular nerve allograft. Synapse;2012:66:52-60

McDermott BM Jr; Asai Y; Baucom JM; Jani SD; Castellanos Y; Gomez G; McClintock JM; Starr CJ; Hudspeth AJ Transgenic labeling of hair cells in the zebrafish acousticolateralis system.Gene expression patterns. 2010;10(2-3):113-8

Kozekov ID; Turesky RJ; Alas GR; Harris CM; Harris TM; Rizzo CJ. Formation of Deoxyguanosine Cross-Links from Calf Thymus DNA Treated with Acrolein and 4-Hydroxy-2-nonenal. Chemical Research in Toxicology.2010;23(11):1701-1713

Stephanie M Cohen, Terrence S Furey, Norman A Doggett, and David G Kaufman. Genome-wide sequence and functional analysis of early replicating DNA in normal human fibroblasts BMC Genomics.2006;7: 301

Dr. Domon, National Agricultural Research Center for Kyushu Okinawa Region, Japan, Extraction of Rush DNA, unpublished personal correspondence, 2004.

Krupey, J., et al, 100,000+ PCRs Possible from 10 ml Blood, poster Biotechniques Symposium, 2003.

Quiniou SM; Katagiri T; Miller NW; Wilson M; Wolters WR; Waldbieser GC Construction and characterization of a BAC library from a gynogenetic channel catfish Ictalurus punctatus. Genetics, selection, evolution : GSE. 2003;35(6):673-83

Charles G. Thornton; Kerry M. MacLellan; Judith R. Stabel; Christine Carothers; Robert H. Whitlock; Selvin Passen. 

 Method to Recovery of Mycobacterium avium subsp. paratuberculosis from Ruminant Tissue Specimens. Journal of Clinical Microbiology.2002;40(5):1783-1790

Charles G. Thornton; Kerry M. MacLellan; Judith R. Stabel; Christine Carothers; Robert H. Whitlock; Selvin Passen Application of the C18-Carboxypropylbetaine Specimen Processing Method to Recovery of Mycobacterium avium subsp. paratuberculosis from Ruminant Tissue SpecimensJournal of Clinical Microbiology.40:5;1783-1790

Applied Biosystems User Bulletin. Subject: Sequencing Large DNA Templates.

J M Kelley; C E Field; M B Craven; D Bocskai; U J Kim; S D Rounsley; M D Adams. High Throughput Direct End Sequencing of BAC Clones. Nucleic Acids Research.1999.15;27(6):1539-1546

Miller, J.M., et al, Polymerase chain reaction identification of Mycobacterium avium in formalin-fixed, paraffin-embedded animal tissues. Journal of veterinary diagnostic investigation.1999.11:436-440

Concentration and purification of beef extract mock eluates from water samples for the detection of enteroviruses, hepatitis A virus, and Norwalk virus by reverse transcription-PCR. Applied and environmental microbiology.1995;61(2):531-7

J D Burton; R N Bamford; C Peters; A J Grant; G Kurys; C K Goldman; J Brennan; E Roessler; T A Waldmann. A lymphokine, provisionally designated interleukin T and produced by a human adult T-cell leukemia line, stimulates T-cell proliferation and the induction of lymphokine-activated killer cellsProceedings of the National Academy of Sciences of the United States of America.1994. 91(11): 4935-4939

DNA Extraction Genetic Diversity as an Indicator of Ecosystem Condition and Sustainability for Regional Assessments of Stream Condition in the Eastern United States

David C. Bruce; Mark O. Mundt; Kim K. McMurry; Linda J. Meincke; Donna L. Robinson; Norman A. Doggett; Larry L. Deaven. BAC Library End Sequencing in Support of Whole Genome AssembliesDOE Joint Genome Institute and Center for Human Genome Studies, Los Alamos National Laboratory

Zoltan S.; Gyimesi; Ilse H.; Stalis; Janice M.; Miller; Charles O Detection of Mycobacterium avium Subspecies avium in Formalin-Fixed, Paraffin-Embedded Tissues of Captive Exotic Birds Using Polymerase Chain Reaction . Thoen Journal of Zoo and Wildlife Medicine.1999;30:3:348-353

Schwab KJ; De Leon R; Sobsey MD. Immunoaffinity concentration and purification of waterborne enteric viruses for detection by reverse transcriptase PCR. Applied and Environmental Microbiology.1996;62(6):2086-94

LA Jaykus, R De Leon and MD Sobsey. A virion concentration method for detection of human enteric viruses in oysters by PCR and oligoprobe hybridization Applied and Environmental Microbiology.1996; 62(6): 2074-2080

Schwab KJ, De Leon R, Sobsey MD Concentration and purification of beef extract mock eluates from water samples for the detection of enteroviruses, hepatitis A virus, and Norwalk virus by reverse transcription-PCR. Applied and Environmental Microbiology.1995; 61(2): 531-537

G.M. Huang; K. Wang; C.L. Kuo; B. Paeper; L. Hood A High-Throughput Plasmid DNA Preparation Method Analytical Biochemtry.1994;15;223(1):35-8

Robert R. Klein, Daryl T. Morishige, Patricia E. Klein, Jianmin Dong; John E. Mullet. High Throughput BAC DNA Isolation for Physical Map Construction of Sorghum. Plant Molecular Biology Reporter.1988;16(4):651-364

 

High-Throughput Genomic DNA Isolation Kit  

 

 
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Biotech Support Group
1 Deer Park Drive, Suite M
Monmouth JCT, New Jersey 08852
sales@biotechsupportgroup.com
Tel:732-274-2866, 1-800-935-0628
Fax: 732-274-2899

 



ProPrep™ is a complete purification system based upon the propietary reagent, ProCipitate™, that has been demonstrated to provide high quality DNA suitable for automated fluorescent sequencing of small to large insert DNA. The ProPrep™ strategy is to remove only the contaminants and leave DNA alone. DNA does not bind, only protein binds.

 

Genomic Sample Preparation Products

 

  

 

Characteristics Of Proprep™

 

ProPrep™ Plasmid & BAC

  • Ideal for mini-prep BAC template preparation from 1.5ml 2xYT cultures.
  • Provides high-quality DNA suitable for automated fluorescent sequencing for small-large insert DNA.
  • Utilizes ProPrep™ strategy which minimizes shearing and improves yield.
  • Adapts easily to robots and automation.
 
 
   

ProPrep™ Omni 

  • Ideal for template DNA preparation and transfection of plasmids, cosmids, and BACs.
  • Provides high-quality DNA suitable for automated fluorescent sequencing for small-large insert DNA.
  • Utilizes ProPrep™ strategy, minimizes shearing and improves yield. Yields to 250ug BAC DNA.
  • Starts with up to 200 & 1000ml cultures.

 


Characteristics Of Procipitate™ 

  • Non-hazardous substitute to phenol/chloroform
  • Removes only the contaminants & leaves DNA alone
  • Improves yield of DNA over alternative bind and elute systems
  • Adaptable to any sample size, and can be automated
  • Key component of the ProPrep™ line of application specific kits

ProCipitate™ is a unique protein removal reagent based upon patented elastomeric polyelectrolytes. The polymer chains of ProCipitate™ are initially extended in a high energy state due to an overall net negative charge. When introduced to protein solutions, the charges are neutralized and the polymer chains collapse to a more favorable energy state; DNA and RNA remain unreacted. ProCipitate™ is non-hazardous and can replace phenol/chloroform with the additional benefits of solid-phase suspensions: adaptability to filtration and automation. It is routinely used for plasmids, cosmids, BACs, and genomic DNA, as well as RNA. ProCipitate™ can also be used to remove Proteinase K and other enzymes. ProCipitate™ provides high quality DNA suitable for automated sequencing, southern blotting, and restriction digestion. ProCipitate™ is available as a suspension reagent and in ProPrep™ kits for specific applications and high-throughput 96 well filter formats.


 

 

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Biotech Support Group | 1 Deer Park Drive, Suite M | Monmouth JCT | NJ | 08852