Home > Courses > Biotechnology > BT734

BT734 : Research Methods in Biotechnology

Course Overview

Course Synopsis

This course is designed to equip students with the multidisciplinary skills and knowledge required in the field of biotechnology. In this course, students will learn the basic skills of the biotechnology workplace, aseptic techniques, preparation of the culture media, isolation of plasmid DNA, quantitation of DNA, polymerase chain reaction, restriction digestion and agarose gel electrophoresis, southern transfer, preparation, purification, and hybridization of probe, transformation, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, western blotting and DNA sequencing. This course will enable the students to get sufficient knowledge about the subject to carry out research and pursue career in the same field.

Course Learning Outcomes

After the successful completion of the course, students will be able to

  • Develop essential research and practical skills in the field of biotechnology.
  • Prepare different types of culture media
  • Isolate and quantify plasmid DNA
  • Perform PCR, restriction digestions, agarose gel electrophoresis, PAGE, Western blotting and DNA sequencing
  • Work independently and take initiative in solving the research problems that may be encountered


Course Calendar

TopicLectureResourcePage
Format of Student Laboratory Records1PPTs
Format of Student Laboratory Records 22PPTs
The Ten Commandments of Record Keeping3PPTs
The Ten Commandments of Record Keeping 24PPTs
Record Keeping and Safety Rules5PPTs
Record Keeping and Safety Rules 26PPTs
Record Keeping and Safety Rules 37PPTs
The Basic Skills of the Biotechnology Workplace: Measuring Volumes8PPTs
The Basic Skills of the Biotechnology Workplace: Measuring Volumes using Pipettes9PPTs
The Basic Skills of the Biotechnology Workplace: Measuring Volumes using MIcropipettes10PPTs
The Basic Skills of the Biotechnology Workplace: Measuring Volumes using MIcropipettes 211PPTs
Basic Skills of the Biotechnology Workplace: Making Solutions12PPTs
Basic Skills of the Biotechnology Workplace: Solutions: Mass/Volume Concentration13PPTs
Basic Skills of the Biotechnology Workplace: Solutions of Differing % Mass/Volume Concentrations14PPTs
Solutions of differing Molar Concentrations15PPTs
Basic Skills of the Biotechnology Workplace: Solutions; Parallel dilution of Existing Stock solution16PPTs
Basic Skills of the Biotechnology Workplace: Solutions; Serial dilution of Existing Stock solution17PPTs
Aseptic Techniques: Introduction18PPTs
Aseptic Techniques: Purpose19PPTs
Aseptic Techniques: Types20PPTs
Aseptic Technique & Establishing Pure Cultures: Introduction21PPTs
Aseptic Technique & Establishing Pure Cultures: Reagents and Supplies22PPTs
Aseptic Technique & Establishing Pure Cultures: Equipment23PPTs
Aseptic Technique & Establishing Pure Cultures: Streak Plate24PPTs
Aseptic Technique & Establishing Pure Cultures:Culture transfer25PPTs
The Growth Curve: Introduction26PPTs
The Growth Curve: Reagents and Supplies27PPTs
The Growth Curve: Equipment28PPTs
The Growth Curve: Procedure29PPTs
The Growth Curve: Cell count using hemocytometer30PPTs
The Growth Curve: Spectrophotometer31PPTs
Assignment No. 1
The Growth Curve: Spectrophotometer Types and Procedure32PPTs
Preparation of the Culture Media: Introduction33PPTs
Preparation of culture Media: Reagents and Supplies34PPTs
Preparation of culture Media: Equipment35PPTs
Preparation of Culture media: Preparation of YEPD (Complex) medium36PPTs
Preparation of Culture Media: Preparation of YNB (Defined medium)37PPTs
DNA Isolation Methods: Preparation of Total Cell DNA38PPTs
Isolation of Plasmid DNA: The Mini-Prep: Introduction39PPTs
Isolation of Plasmid DNA: The Mini-Prep: Reagents and Supplies40PPTs
Isolation of Plasmid DNA: The Mini-Prep: Equipment41PPTs
Isolation of Plasmid DNA: The Mini-Prep: procedure42PPTs
Purification of DNA: Introduction43PPTs
Purification of Plasmid DNA by solvent extraction: Reagents ,supplies and equipment44PPTs
Purification of Plasmid DNA by solvent extraction: Procedure45PPTs
Concentration of Plasmid DNA: Reagents , supplies and equipment46PPTs
Concentration of Plasmid DNA: Procedure47PPTs
Quantitation of DNA: Reagents ,supplies and equipments48PPTs
Quantitation of DNA: Procedure (UV absorption)49PPTs
Quantitation of DNA: Procedure (Etbr Flourescence)50PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography): Introduction51PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography):Ion Exchange Chromatography52PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography):Reagents\Supplies53PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography): Equipments54PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography): Procedure55PPTs
Large Scale Isolation of Plasmid DNA (Column chromatography):CsCl Density gradient56PPTs
Amplification of a gene by (PCR): Introduction57PPTs
Amplification of a gene by (PCR): Parameters that affect PCR58PPTs
Amplification of Gene Fragment by PCR: Thermostable DNA Polymerases59PPTs
Amplification of Gene Fragment by PCR: Programing a PCR60PPTs
Amplification of Gene Fragment by PCR: Selecting PCR Primers61PPTs
Amplification of Gene Fragment by PCR: Properties of Oligonucleotides and efficiency of amplification62PPTs
Assignment No. 2
Amplification of Gene Fragment by PCR: Calculating melting temperature ™63PPTs
Amplification of Gene Fragment by PCR: Contamination in PCR64PPTs
Amplification of Gene Fragment by PCR: Reagents and Supplies65PPTs
Amplification of Gene Fragment by PCR: Procedure66PPTs
Amplification of Gene Fragment by PCR: Trouble shooting the amplification67PPTs
Amplification of Gene Fragment by PCR: Troule shooting major problems that occur in PCR68PPTs
Amplification of Gene Fragment by PCR: Purification of PCR Products in Preparation for Cloning69PPTs
Purification of PCR products: Reagents, Supplies and Equipment70PPTs
Purification of PCR products: Procedure71PPTs
Removal of Primers and Excess dNTPs from Amplified DNA by Ultrafication: Introduction72PPTs
Removal of Primers and Excess dNTPs from Amplified DNA by Ultrafication: Reagents supplies and Equipment73PPTs
Removal of Primers and Excess dNTPs from Amplified DNA by Ultrafication: Procedure74PPTs
Amplification of Gene fragment by PCR: Types of PCR75PPTs
Gel Electrophoresis: Introduction76PPTs
Agarose Gel electrophoresis: Agarose77PPTs
Agarose Gel electrophoresis: The rate of migration of DNA through agarose gels78PPTs
Agarose Gel electrophoresis: Electrophoresis Buffers79PPTs
Gel electrophoresis: Gel Loading Buffer80PPTs
Agarose Gel electrophoresis: Reagents ,Supplies and Equipments81PPTs
Agarose Gel electrophoresis: Procedure82PPTs
Agarose Gel Electrophoresis: Detection of DNA in Agarose gel83PPTs
Agarose Gel Electrophoresis: Staining with Sybr Gold84PPTs
Agarose Gel Electrophoresis: Photography of DNA in Gel85PPTs
Agarose Gel Electrophoresis: Recovery of DNA from Low-melting-temperature Agarose Gels Organic Extraction86PPTs
Agarose Gel Electrophoresis: Reagents and supplies Recovery of DNA from Low-melting-temperature Agarose Gels Organic Extraction87PPTs
Agarose Gel Electrophoresis: Procedure Reagents and supplies Recovery of DNA from Low-melting-temperature Agarose Gels Organic Extraction88PPTs
Southern Hybridization: Introduction89PPTs
Southern Hybridization: Methods to transfer DNA from Agarose gel to solid support90PPTs
Southern Hybridization: Membranes used for Southern and Northern blotting91PPTs
Southern Hybridization: Capillary transfer method Southern Blot92PPTs
Southern Hybridization: Restriction digestion of DNA for southern93PPTs
Southern Hybridization: Restriction digestion [Problems and Solutions]94PPTs
Southern Hybridization: Reagents and Supplies95PPTs
Southern Hybridization:Procedure [Digestion & Electrophoresis]96PPTs
Southern Hybridization: Preparation of gel for transfer97PPTs
Southern Hybridization:Preparation of membrane for transfer98PPTs
southern hybridization:Apparatus assembly for DNA transfer99PPTs
Southern Hybridization:Fixation of DNA to membrane100PPTs
Southern Hybridization: Southern blot: hybridization of radiolabeled probe to nucleic acids101PPTs
Probe hybridization to Nucleic Acids:Reagents and Supplies102PPTs
Probe hybridization to Nucleic Acids:Hybridization solutions for different buffers103PPTs
Probe hybridization to Nucleic Acids:Salmon Sperm DNA104PPTs
Probe hybridization to Nucleic Acids:Special Equipment105PPTs
Probe hybridization to Nucleic Acids:Procedure106PPTs
Probe hybridization to Nucleic Acids: Stripping Probes from Membranes107PPTs
Southern Hybridization:Method: Stripping Probes from Membranes108PPTs
Southern Hybridization:Method: Stripping Probes (Nylon Membrane)109PPTs
Southern Hybridization: Probe synthesis:Nick Translation radioactive labeling110PPTs
Southern Hybridization: Probe synthesis:End filling radioactive labeling111PPTs
Southern Hybridization: Probe synthesis:Random primed radioactive labeling112PPTs
Random Priming; Radiolabeling of DNA:Introduction113PPTs
Random Priming; Radiolabeling of DNA:Reagents and Supplies114PPTs
Random Priming; Radiolabeling of DNA:Procedure115PPTs
Purification of RNA from Cells & Tissues:Introduction116PPTs
Purification of RNA from Cells & Tissues:Reagent & Supplies (Acid Phenol-Guanidinium Thiocyanate-Chloroform Extraction117PPTs
Purification of RNA from Cells & Tissues:Procedure118PPTs
Purification of RNA from Cells & Tissues:Procedure_2119PPTs
Purification of RNA from Cells & Tissues:Procedure_3120PPTs
Purification of RNA from Cells & Tissues:Procedure [Extraction of RNA from Homogenates121PPTs
Purification of RNA from Cells & Tissues:Storage of RNA122PPTs
Simultaneous Preparation of DNA, RNA, & Protein:Introduction123PPTs
Simultaneous Preparation of DNA, RNA, & Protein:Reagent & Supplies124PPTs
Simultaneous Preparation of DNA, RNA, & Protein:Procedure125PPTs
Special Assignment (in lieu of Midterm Examination)
Simultaneous Preparation of DNA, RNA, & Protein:Procedure_2126PPTs
Simultaneous Preparation of DNA, RNA, & Protein:Procedure_3127PPTs
Simultaneous Preparation of DNA, RNA, & Protein:Procedure_4128PPTs
Purification of Poly A RNA:Introduction129PPTs
Purification of Poly A RNA:Reagents & Supplies130PPTs
Purification of Poly A RNA:Procedure131PPTs
Purification of Poly A RNA:Further Purification of Eluted RNA132PPTs
Northern Hybridization:Introduction133PPTs
Electrophoresis [Glyoxylated RNA]:Introduction134PPTs
Electrophoresis [Glyoxylated RNA]:Reagents and supplies135PPTs
Electrophoresis [Glyoxylated RNA]:Procedure136PPTs
Quality of RNA:How to check quality of RNA preparation137PPTs
Gel Electrophoresis [Formaldehyde]:Introduction138PPTs
Gel Electrophoresis [Formaldehyde]:Reagents and supplies139PPTs
Gel Electrophoresis [Formaldehyde]:Procedure140PPTs
Transfer and Fixation of Denatured RNA to membrane Introduction (1)141PPTs
Reagents and Supplies for transfer and fixation of denatured RNA142PPTs
Procedure Transfer and Fixation of denatured RNA143PPTs
Procedure Assembly of the transfer system144PPTs
Procedure Staining of RNA and fixation of RNA on membrane145PPTs
Northern Blotting introduction146PPTs
Reagents and Supplies for Northern blotting147PPTs
Northern blotting: Procedure148PPTs
Protein assay Introduction Biuret149PPTs
Reagents and Supplies for Biuret150PPTs
Procedure biuret assay      Current Lesson151PPTs
Assignment No. 3
Protein assay Introduction Lowry152PPTs
Reagents and Supplies for Lowry assay153PPTs
Procedure Lowry assay154PPTs
Bradford protein assay155PPTs
Reagents and Supplies Bradford protein assay156PPTs
Procedure Bradford assay157PPTs
Protein Purification from Inclusion bodies: Introduction158PPTs
Protein Purification from Inclusion bodies: Reagent and Supplies159PPTs
Purification of Proteins: Procedure (Preparation of cell Extract)160PPTs
Purification of Proteins: Procedure (Purification and washing of inclusion bodies)161PPTs
Purification of Proteins: Procedure (Purification and washing of inclusion bodies) 1162PPTs
Purification of Proteins: Procedure (Solubilization of Inclusion Bodies)163PPTs
Protein Isolation from Animal Tissues: Introduction164PPTs
Protein Purification from Animal Tissues: Reagents and Supplies165PPTs
Protein Extraction from Animal Tissues: Procedure166PPTs
Precipitation with Ammonium Sulfate: Introduction167PPTs
Precipitation with Ammonium sulfate:Reagents & Supplies168PPTs
Precipitation with Ammonium sulfate:Procedure169PPTs
SDS-PAGE:Introduction170PPTs
SDS-PAGE:Reagents & Supplies171PPTs
SDS-PAGE:Reagents & Supplies 2172PPTs
SDS-PAGE:Procedure173PPTs
SDS-PAGE:Procedure 2174PPTs
SDS-PAGE:Procedure Staining175PPTs
Western Blotting:Introduction176PPTs
Transfer of protein to membrane:Procedure177PPTs
Transfer of protein to membrane:Procedure2178PPTs
PROBE FOR WESTERN BLOT:Reagents & Supplies179PPTs
Immunoblotting:Procedure180PPTs
ELISA:Introduction181PPTs
ELISA:Types of ELISA182PPTs
ELISA:Sandwich ELISA183PPTs
ELISA:COMPETITION OR INHIBITION ELISA184PPTs
ELISA:Reagents & Supplies185PPTs
ELISA:Procedure186PPTs
ELISA RESULTS:Results187PPTs
Microarray:Introduction188PPTs
MICROARRAY:Types of Microarray189PPTs
Microarray:Reagents & Supplies190PPTs
Microarray:Procedure191PPTs
Transgenic Mouse (TM) Preparation:Introduction192PPTs
Transgenic Mouse (TM) Preparation:Reagents & Supplies193PPTs
Transgenic Mouse (TM) Preparation:Animals and Drugs194PPTs
Transgenic Mouse (TM) Preparation:Procedure Egg collection195PPTs
Transgenic Mouse (TM) Preparation:Procedure Egg collection 2196PPTs
Transgenic Mouse (TM) Preparation:Procedure;Injection of eggs197PPTs
Transgenic Mouse (TM) Preparation:Procedure;Implanting Eggs in Foster Mothers198PPTs
DNA Sequencing:Introduction199PPTs
DNA Sequencing:Types of DNA Sequencing200PPTs
DNA Sequencing:The Chemical Method of DNA Sequencing201PPTs
Preparing template for ddNTP Sequencing:Introduction202PPTs
Preparing template for ddNTP Sequencing:Reagents & Supplies203PPTs
Preparing template for ddNTP Sequencing:Procedure204PPTs
ddNTP Sequencing; Taq DNA Polymerase:Introduction205PPTs
ddNTP Sequencing; Taq DNA Polymerase:Reagents & Supplies206PPTs
Preparing template for ddNTP Sequencing:Procedure 2207PPTs
ddNTP sequencing using Sequenase:Introduction208PPTs
ddNTP Sequencing; Sequenase:Reagents & Supplies209PPTs
Preparing template for ddNTP Sequencing:Procedure(Sequenase)210PPTs
ddNTP sequencing; Klenow fragment:Introduction211PPTs
ddNTP Sequencing; Taq DNA Polymerase:Reagents & Supplies 2212PPTs
ddNTP Sequencing; Klenow:Procedure213PPTs
ddNTP (Cycle) Sequencing:Introduction(Using PCRand End- labeled Primers)214PPTs
ddNTP (Cycle) Sequencing:Introduction;Advantage215PPTs
ddNTP (Cycle) Sequencing:Reagents & Supplies216PPTs
ddNTP Cycle Sequencing:Procedure217PPTs
Chemical Sequencing:Introduction218PPTs
Chemical Sequencing:Reagents & Supplies219PPTs
Chemical Sequencing:Procedure220PPTs
Cell Culture:Introduction221PPTs
Cell Lines:Introduction Cell Lines222PPTs
Cell Culture types:Introduction223PPTs
When to Subculture:Introduction224PPTs
Cell Culture:Equipment225PPTs
Cell Culture:Laboratory226PPTs
Cell Culture:Cell culture Hood Layout227PPTs
Cell Culture:Laboratory III228PPTs
Cell Culture:Laboratory IV229PPTs
Trypsinizing and Subculturing:Trypsinizing and subculturing cells from a monolayer230PPTs
Trypsinizing and Subculturing:Procedure:Trypsinizing and subculturing cells from a monolayer231PPTs
Cell Passage:Passaging cells in Suspension Culture232PPTs
Freezing Human Cell:Freezing Human cells grown in monolayer cultures233PPTs
Suspension Culture:FREEZING CELLS GROWN IN SUSPENSION CULTURE234PPTs
Thaw and Recovery:Thawing and recovering human cells235PPTs
Thaw and Recovery:Procedure236PPTs
HEMOCYTOMETER AND TRYPAN BLUE STAINING:DETERMINING CELL NUMBER AND VIABILITY237PPTs
Cell Transport:PREPARING CELLS FOR TRANSPORT238PPTs
siRNA [Small interfering RNA]:Introduction239PPTs
siRNA: Where do they come from?240PPTs
siRNA:siRNA designing241PPTs
siRNA:Mechanism242PPTs
siRNA:Applications243PPTs
Final term Exam
 
 
Back to Top