Panoramic Parvovirus Strain Discovery Data Chain

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ParvoDB ID PV005458 GenBank Accession AY386330
Subfamily Parvovirinae Genus Erythroparvovirus
Species Erythroparvovirus primate1 Sequence Length 5596
Definition B19 virus isolate J35, complete genome. Submission Date 2003-09-09
Strain Name J35 Sampling Date
Sampling Country None Location None
Submitted By Brown,K.E., Zhi,N., Zadori,Z. and Tijssen,P. Submitting Institution Hematology Branch, National Heart Lung and Blood Institute, 9000 Rockville Pike, Bethesda, MD 20892, USA
Original Host human Sample Type None
Standardized Host Name Homo sapiens Host Tax Rank species
Standardized Host Class Mammalia Standardized Host Order Primates
Environmental Origin - N/A - Reagent Origin - N/A -
Number Reagent/Consumables Purpose PMID/Url Reference
1 B19 DNA (J35 isolate) Source of viral genetic material for cloning and transfection purposes. 14972543 View Related Publication
2 pCR-Blunt II TOPO vector Used for blunt ligation of PCR amplified B19 coding region.
3 Top10 cells Transformation host cells for holding B19 coding region clones.
4 32P-random-primed B19 probe Used for hybridization to screen colonies containing B19 inserts.
5 Sure2 cells Transformation host cells used for holding and cloning full-length B19 genome.
6 HF-2 polymerase kit For high-fidelity long PCR amplification.
7 pProEX HTb vector Used for cloning digested fragments of B19 DNA.
8 AMAXA Cell Line Nucleofector kit R Optimization and successful transfection of plasmid DNA into UT7/Epo-S1 cells.
9 Isocove's Modified Dulbecco's Medium (IMDM) Culture medium for UT7/Epo-S1 cell line.
10 Fetal calf serum (FCS) Supplement for cell culture medium.
11 Recombinant human erythropoietin (Epo, AMGEN) Supplement for inducing UT7/Epo-S1 cell growth.
12 RNA STAT60 For RNA extraction from cells.
13 RT-PCR primers (B19-1, B19-9, B19-2255, B19-2543) For PCR amplification of specific regions of B19 transcripts.
14 SuperScript II Reverse transcriptase for converting RNA to cDNA.
15 Monoclonal antibody 521-5D Primary antibody specific to B19 capsid proteins for immunofluorescence assay.
16 Fluorescein isothiocyanate-labeled goat anti-mouse IgG Secondary antibody used in immunofluorescence assay.
17 DNAse I For removing residual genomic DNA from RNA preparations.
18 HindIII, BssHII, SalI restriction enzymes For mapping and constructing recombinant plasmids.
19 Hirt method reagents (NaCl, Tris-HCl, SDS, EDTA, Proteinase K) For separating high- and low-molecular weight DNA.
20 Ecl136II, XhoI, MscI, XbaI restriction enzymes For eliminating undesired restriction sites and construction of variant plasmid.
21 BamHI, EcoRI restriction enzymes For detecting replication intermediates in Southern blot analysis.
22 RNase (Roche) Treating cell lysates to degrade RNA post-clarification.
23 Monoclonal antibodies A and E Used in neutralization assays to confirm the infectivity mediated by newly synthesized B19 virions.