A point mutation in the iron-responsive element of the L-ferritin gene: DNA analysis

At 22 years of age, the patient underwent weekly 500 mL phlebotomy treatments and dropped her hemoglobin level from 153 g/L to 84 g/L over one month. She subsequently developed symptomatic cataracts and, given a positive family history of cataracts, the diagnosis of HHCS was made and she underwent surgical treatment.

Genomic DNA was extracted from peripheral blood leukocytes of eight individuals in the pedigree, four of whom were affected by cataracts. The 5′ region of the L-ferritin gene was amplified by polymerase chain reaction (PCR) using flanking primers (forward 5′-TCCTTGCCACCGCAGATTG-3′; reverse 5′-TTGGCAAGAAGGAGCTAACC-3′ . PCR reactions were made up to a total volume of 50 pL containing 25 pmol of each primer, 1 U Taq polymerase and 2.5 pmol magnesium chloride. PCR reactions were carried out under the following conditions using the PTC-100 Thermocycler: denaturation at 94°C for 30 s, annealing at 54°C for 30 s and extension at 72°C for 90 s. Nested PCR was performed using the same primers to yield a 287-base pair product. PCR products were gel purified and DNA extracted using QIAquick Gel Extraction Kit (QIAGEN Inc, Canada). DNA was sent for direct sequencing at the Robarts Research Institute (London, Ontario) sequencing laboratory. All you need to discover how safe and advantageous it can be to purchase buy generic Tavist online over the internet is visit the pharmacy suggested and enjoy your shopping experience as well as your treatment.


Category: L-ferritin gene

Tags: Cataracts, Hyperferritinemia, Iron-responsive element, L-ferritin

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