Podcast
Questions and Answers
What is the primary function of the Photo-to-PDF One Click Converter?
What is the primary function of the Photo-to-PDF One Click Converter?
Which of the following features might NOT be expected from a converter application?
Which of the following features might NOT be expected from a converter application?
Where can the Photo-to-PDF One Click Converter be downloaded?
Where can the Photo-to-PDF One Click Converter be downloaded?
Which statement is most accurate about the usage of the Photo-to-PDF converter?
Which statement is most accurate about the usage of the Photo-to-PDF converter?
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Which aspect of the Photo-to-PDF converter might attract users?
Which aspect of the Photo-to-PDF converter might attract users?
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What is a likely limitation of the Photo-to-PDF converter?
What is a likely limitation of the Photo-to-PDF converter?
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For what primary purpose would someone use a converter like Photo-to-PDF?
For what primary purpose would someone use a converter like Photo-to-PDF?
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Which of the following would be a common user expectation when using a converter app?
Which of the following would be a common user expectation when using a converter app?
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What type of output can users expect after utilizing the converter successfully?
What type of output can users expect after utilizing the converter successfully?
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Which of the following is a likely advantage of using a one-click converter like this?
Which of the following is a likely advantage of using a one-click converter like this?
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Study Notes
Artificial Chromosome Vectors
- DNA molecules assembled in vitro
- Used to clone large DNA fragments
- Used for genome sequencing
Characteristics
- ORI
- MCS
- Selectable Marker
Types
-
BAC (Bacterial Artificial Chromosome)
- Engineered DNA molecules used to clone DNA in bacterial cells
- Primarily used for DNA sequencing
- DNA fragment size: 150-350kb to 1-3 Lakh bp
- Natural plasmid (E. coli plasmid vector):
- Copy no: 1-2 copies/cell
- Used to study inherited diseases
- Mostly inserted into bacteria through gene gun/electroporation
- Components: origin of replication, copy number regulation, selectable markers (antibiotic resistance genes), and promoters
-
YAC (Yeast Artificial Chromosome):
- Shuttle vectors
- Small & large in size when amplified
- Components: centromere, telomeres, and ARS (autonomous replication sequence)
Lambda Vector
- Carrying capacity: 10kb
- Virus infects and injects viral DNA
- Temperate phage (infects E.coli)
- Replicates via the lytic or lysogenic cycle
- Releases 100+ phages
- Single linear DNA of 50kb
- Contains cos sites at both ends with cohesive ends
Insertion Vector/Replacement Vector
- Structures include BamHI, BarnTI, L/R, Stuffer, fragments
- Invitro construction:
- Cut source DNA with BamHI
- Isolate 15-20kb DNA fragments
- Ligate with DNA ligase
- Add empty bacteriophage tail
- Pack DNA into phage vectors
In vitro Packaging
- Pre-heads are made (gene encoded protein)
- Inserting DNA into pre-heads
- Joining & forming proteins/DNA into a new structure
Agrobacterium tumefaciens-based Vectors
- Tumor-inducing size >200kb
- T-DNA transferred to Ti plasmid
- 25 bp, 23kb, 206 bp (borders)
Co-integrate Vector
- By homologous recombination
- Engineered Ti plasmids with T-DNA inserts
Binary Vector
- Mini-Ti plasmid
- LB & RB
- Foreign DNA
- Instead of T-DNA, other genes are used(e.g., helper Ti plasmid)
Cloning Vector/Expression Vector
- Cloning vector: replicates in host cells (just copies)
- Expression vector: makes proteins in host cells
- Used to obtain multiple clones of specific genes (e.g., from SM, Ori, MCS)
- Expression vectors: used to obtain genes/products like RNA or proteins
- Enhances (e.g., promoter, start/stop codon, terminator)
- Plasmid vector, naturally occurring, smallest & most used, 1-250kb, e.g., PBR322, PUC18/19
Other PCR Methods
- Nested PCR
- Hot start PCR
- Real-time PCR
- Multiplex PCR
- Colony PCR
- Asymmetric PCR(used to amplify one strand)
Other Blotting Techniques
- Southern blotting: detects specific DNA sequences
- Northern blotting: detects specific RNA sequences
- Western blotting: detects specific protein sequences
Other Methods
- Nick translation
- Polymerase Chain Reaction (PCR)
Capsular Material/Chromosomal Material
- Used in gene gun method to deliver a desired gene
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