GeneQuery™ HLA-C PCR+Sanger SBT Typing Kit

The human leukocyte antigen (HLA) complex is a gene complex located on the short arm of human chromosome 6. The HLA complex is the human version of the major histocompatibility complex (MHC) which helps the immune system distinguish host cells from invaders. It is the most polymorphic locus in the human genome, and has been implicated in many pathologies including autoimmunity disorders and cancer.

GeneQuery™ HLA-C PCR+Sanger SBT Typing Kit (SBT-C) utilizes the Sequence-based Typing (SBT) method, which is the gold standard for HLA typing. This method involves PCR amplification of HLA genes, followed by Sanger sequencing of the amplicons. The acquired sequences are aligned to an HLA allele database using the SOAPTyping software (BGI Genomics) to obtain the 4-digit genotypes of the samples. The two haplotypes from both parents can be typed simultaneously using this kit and the SOAPTyping software.

SBT-C kit contains a ScienCell proprietary HLA-C amplification primer set (C_ampF primer mix and C_ampR primer mix; Cat #GK893a) that can amplify a region in the human genome that covers exons 2-4 of HLA-C gene, and 6 sequencing primer mixes (C_2F, C_2R, C_3F, C_3R, C_4F, and C_4R; Cat #s GK893b-g) that can be used in the following Sanger sequencing of the PCR product. Please refer to Figure 2 for the binding location of the primers on the human genome. Each GeneQuery™ HLA typing kit contains a human Genomic DNA Reference (GDR) sample with known HLA alleles (Cat #GK893r). The GDR can be used to monitor the quality of the reagents and the instruments. The HLA-C alleles in GK893r are HLA-C*07:01 and HLA-C*12:03.

Primer binding locations on the human genome.

Additional Materials Required (Materials Not Included in Kit)

Component	Recommended
genomic DNA template	Customers’ samples
PCR purification kit	SpeeDNA PCR Purification Kit (ScienCell, Cat #MB6948)
Sanger sequencing kit
PCR plates or tubes

Catalog No.	GK893
Country of Manufacture	United States
Product Code	SBT-C
Size/Quantity	40 samples
Product Use	SBT-C is for research use only. It is not approved for human or animal use, or for application in clinical or in vitro diagnostic procedures.
Storage	Primers & EmeraldNStart HiFi Marathon PCR Master Mix at -20°C in a manual defrost freezer, nuclease-free H2O at 4°C. Once the EmeraldNStart HiFi Marathon PCR Master Mix is thawed, store at 4°C, do not refreeze. It's stable at 4°C for up to 6 months.
Shipping Info	SBT-C is shipped on dry ice.

Product Sheets

GK893

Certificates of Analysis

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Cat #	Component	Quantity	Storage
GK893a	HLA-C amplification primer set (C_ampF and C_ampR)	400 µL	-20°C
GK893b	C_2F sequencing primer mix, 10 µM	200 μL	-20°C
GK893c	C_2R sequencing primer mix, 10 µM	200 μL	-20°C
GK893d	C_3F sequencing primer mix, 10 µM	200 μL	-20°C
GK893e	C_3R sequencing primer mix, 10 µM	200 μL	-20°C
GK893f	C_4F sequencing primer mix, 10 µM	200 µL	-20°C
GK893g	C_4R sequencing primer mix, 10 µM	200 μL	-20°C
GK893h	Nuclease-free H₂O	4 mL	4°C
GK893r	Human genomic DNA reference (GDR), ~50 ng/µL (HLA-C07:01 and HLA-C12:03)	10 μL	-20°C
MB6038-1	2X EmeraldNStart HiFi Marathon PCR Master Mix, 1 mL	2 vials	-20°C

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