Applied Biosystems™ HIV-1 Genotyping Kit with Integrase

• Designed to genotype the protease, reverse transcriptase (RT) and integrase (IN) regions of the HIV-1 pol gene
• The genotyping result encompasses codons 6-99 in the protease region and codons 1-251 in the RT region of the gene
• HIV-1 genomic mutations in these regions confer resistance to anti-retroviral drugs that act through the inhibition of the protease and reverse transcriptase
• The kit enables reliable genotyping of the genetically diverse HIV-1 virus to investigate Drug Resistance Mutations (DRMs) to protease inhibitors (PI) , nucleoside reverse-transcriptase inhibitors (NRTI), non-nucleoside reverse-transcriptase inhibitors (NNRTI), and integrase inhibitors (INSTI) from HIV-1 RNA extracted from EDTA plasma and dried blood spot samples
• This product employs assays for HIV-1 genotyping licensed from the U.S. Centers for Disease Control and Prevention
• Kit contains ready-to-use master mix, enzyme, and positive and negative controls to cover the three phases of the workflow: RT-PCR, nested PCR, and bidirectional cycle sequencing steps to process 48 test samples
• Starting with viral RNA extracted from an EDTA plasma or dried blood spot sample, RT-PCR is conducted to generate PR/RT and IN PCR products as DNA template
• After confirming the band size of the PCR amplicons, the Cycle Sequencing module is used to sequence the DNA templates utilizing Sanger sequencing technology
• The sequencing reactions are first purified, then sequenced using POP-7 on a designated capillary electrophoresis system
• The sequencing traces are reviewed for quality, after which the user exports the AB1 file for use in downstream analysis and reporting software a mutation profile through a curated database

Includes

• RNA Positive Control, 2 x 100 μL
• HIV RNA Negative Control, 2 x 100 μL
• RT-PCR Master Mix, PR/RT, 2 x 1.055 mL
• RT-PCR Master Mix, IN, 2 x 1.055 mL
• Nested-PCR Master Mix, PR/RT, 2 x 1.285 mL
• Nested-PCR Master Mix, IN, 2 x 1,285 μL
• SuperScript™ III One-Step RT-PCR with Platinum™ Taq High Fidelity Enzyme, 2 x 54 μL
• AmpliTaq Gold™ LD DNA Polymerase, 1 x 54 μL
• Sequencing vector, 2 x 80 μL
• HIV Sequencing Mix R12, 2 x 435 μL
• HIV Sequencing Mix R11, 2 x 435 μL
• HIV Sequencing Mix F12, 2 x 435 μL
• HIV Sequencing Mix F11, 2 x 435 μL
• HIV Sequencing Mix R3, 2 x 435 μL
• HIV Sequencing Mix R2, 2 x 435 μL
• HIV Sequencing Mix R1, 2 x 435 μL
• HIV Sequencing Mix F3, 2 x 435 μL
• HIV Sequencing Mix F2, 2 x 435 μL
• HIV Sequencing Mix F1, 2 x 435 μL

Store all components at -15°C to -25°C