Group b

Cloning Simulation Project 2003 GTB 204 - Molecular Biology Techniques

Abstract ,[object Object],[object Object]

Introduction ,[object Object],[object Object],[object Object],[object Object]

[object Object],[object Object],[object Object],[object Object]

Objective ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Step1: Hunting for the gene ,[object Object],[object Object],[object Object],[object Object]

Step 2: Determining Amino Acid Sequence & Molecular Weight of Erythropoietin Gene

[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Step 3: Looking for Open Reading Frame (ORF) ,[object Object],[object Object],[object Object],[object Object],[object Object]

ORF Finder (Open Reading Frame Finder)

Step 4: Looking for the Restriction Enzyme Site ,[object Object],[object Object],[object Object],[object Object]

Step 5: Comparing the Gene Sequence with other similar Gene Sequences by BLAST Analysis and Multiple DNA Sequence Alignment ,[object Object]

BLAST (Basic Local Alignment Search Tool) ,[object Object]

Multiple DNA Sequence Alignment (CLUSTALW) ,[object Object],[object Object],[object Object]

Step 6: Designing primer for PCR Amplification and screening ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Calculation of the Annealing Temperature ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Step 7: Designing the Vector ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Step 8: PCR ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

PCR cont. ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

Step 9: TOPO TA Cloning ,[object Object],[object Object],[object Object],[object Object]

TOPO TA Cloning cont. ,[object Object]

TRANSFORMATION INTO AN E.COLI. ,[object Object],[object Object],[object Object],[object Object]

[object Object],[object Object],[object Object],PROCEDURES

VERIFICATION OF SUCCESSFUL TRANSFORMATION ,[object Object],[object Object],[object Object],[object Object]

Step 11: Screening of Recombinant Clone through PCR Method

PROTEIN PURIFICATION ,[object Object],[object Object]

PROTEIN PURIFICATION nickel metal affinity chromatography method

PROTEIN PURIFICATION ,[object Object],[object Object],[object Object],[object Object],[object Object]

[object Object],[object Object],[object Object],[object Object],[object Object],PROTEIN PURIFICATION

SDS PAGE ,[object Object],[object Object],[object Object]

SDS PAGE ,[object Object],[object Object]

SDS PAGE-cont; ,[object Object],[object Object]

COMMERCIAL VALUES ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

References ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]

THANK YOU SPACIBA! ARIGATO! GRACIAS!

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