Sscp Conditions Optimization: Optimization Of Single Strand Conformational Polymorphism (Sscp)Conditions For Exon 4-12 Of Ldlr Gene

 SSCP Conditions Optimization: Optimization of Single Strand Conformational Polymorphism (SSCP)Conditions for Exon 4-12 of LDLR Gene Text fb2 book

Single strand conformational polymorphism (SSCP) is a technique which is used to detect point mutation in genes. The purpose was to optimize the condition of SSCP for efficient and sensitive mutation detection in Exon 4, 5, 6, 7, 8, 9, 10, 11 and 12. All these Exon were amplified and run on agarose gel to check strong and specific amplification. These samples were then subjected for vertical elect...

Paperback: 68 pages
Publisher: LAP LAMBERT Academic Publishing (November 10, 2010)
Language: English
ISBN-10: 9783843372978
ISBN-13: 978-3843372978
ASIN: 3843372977
Product Dimensions: 5.9 x 0.2 x 8.7 inches
Format: PDF ePub Text TXT fb2 book

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for SSCP using different conditions of gel concentration, time, temperature and voltage to get maximum separation between single strands. Denaturation was carried out by mixing the samples with denaturant reagents. The length of nucleotide of Exon 4 was much longer than normal, so we amplified the exon 4 into two small parts named as 4a and 4b respectively after its bifurcation. The denaturation reagent consist of 50% 100mM NaOH and 50% Formamide loading buffer, 5?l-10?l sample was loaded in each case. In all the optimized Exons the denaturation was performed at 95°C for 7 minutes in PCR machine. This optimization will prove a valuable help in further study of the Exons of the same gene and other genes for Point Mutation Detection.