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1
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Virion DNA
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Purified as template for PCR reactions.
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8774690
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View Related Publication
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2
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PCR Primers (NS forward, NS reverse, VP1 forward, VP2 forward, VP1/2 reverse)
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Used for amplifying specific DNA regions of parvovirus B19 genome.
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3
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Deoxynucleoside triphosphates (0.25 mM)
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Included in PCR reactions to facilitate DNA synthesis.
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4
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PCR Buffer (100 mM Tris-HCl, pH 8.8 at 25°C, 500 mM KCl, 15 mM MgCl2, 1% Triton X-100)
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Provides optimal conditions for Taq polymerase activity during PCR.
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5
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Taq Polymerase (2.5 units)
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Facilitates the polymerase chain reaction to amplify DNA.
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6
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Geneclean kit (Bio 101 Inc.)
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Used for purifying PCR products.
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7
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pGEM-T vector system (Promega)
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Vector system for cloning VP2 PCR product.
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8
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pCR-II using the TA cloning kit (Invitrogen)
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Vector system for cloning VP1 and NS PCR products.
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9
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Baculovirus transfer vector (pVL1392)
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Used to create recombinant baculovirus containing the NS gene.
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10
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T4 DNA Ligase
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Used for ligating DNA fragments into vectors.
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11
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HB101 competent cells
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E. coli cells used for transformation with plasmid DNA.
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12
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CsCl gradient
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Used for purifying large-scale plasmid DNA preparations.
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13
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Sf9 cells (PharMingen)
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Insect cells used for baculovirus infections and recombinant protein expression.
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14
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Wild type Autographica californica nuclear polyhedrosis virus (AcNPV; Pharmingen)
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Used along with recombinant baculovirus to infect Sf9 cells.
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15
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Grace's supplemented insect medium (Gibco BRL)
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Nutrient medium for growing Sf9 cells.
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16
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Fetal calf serum
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Supplement in cell culture media.
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17
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Gentamycin (50 μg/ml)
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Antibiotic added to cell culture media.
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18
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Calcium phosphate
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Used for transfection of Sf9 cells.
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19
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Phosphate buffered saline (PBS)
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Buffer used in various stages of cell processing and Western blot.
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20
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Tween-20
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Detergent used in PBS for washing and blocking steps in Western blot.
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21
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Non-fat dried milk (Marvel)
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Blocking agent in PBS-Tween solution for Western blots.
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22
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Horse radish peroxidase-conjugated anti-human IgG
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Secondary antibody used in Western blotting.
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23
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3',3'-diaminobenzidine (DAB; Sigma)
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Substrate used for colorimetric detection in Western blotting.
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