RR4 DNA cloning, gene expression and genome

How do we sequence DNA with PCR? (illumina sequencing)

Associating a color to a nucleotide, throwing all nucleotides on the sequencing board, wiping the nucleotides that did not stick and checking what was the change in color for each strand being sequenced, and repeat until finished

What is nanopore sequencing?

New generation sequencing, uses pores. Pores 1nm wide to allow single-strand of DNA to pass through. Potential difference across the membrane = ion flow through the pores. As DNA passes through the pore, change in current flow produces a "squiggle". "Squiggle" decoded by base-calling software. Gives long sequence reads

What are plasmids?

Plasmids are small loops of extra DNA that aren't part of the chromosome. Huge diversity of plasmids in nature

What are restriction endonucleases?

Enzymes which recognize specific sequences of DNA and cut the doubled-stranded molecule into two pieces. Each one recognizes a different site

What is the polylinker region on a plasmi?

Its polylinker region is composed of several restriction enzyme recognition sites, that have been engineered into a single region.

What conditions permit the insertion of plasmids in bacteria by humans

CaCl2 and Heat pulse

What are DNA libraries?

Permanent collections of genes.
Divided in two types :
Genomic libraries (stores chromosomal DNA)
cDNA libraries (stores mRNAs that can be reverse transcribed in cDNAs)

What is reverse transcription?

Going from RNA to cDNA using reverse transcriptase

What are some Molecular Genetic Techniques (what uses can you make of you cDNA)

Microarray and in situ hybridization techniques reveal mRNA expression, co-regulation , and localization

Recombinant DNA expression vectors enable regulated expression of exogenous genes and production of proteins

True or False : Cloned genes can be expressed in animal cells?

Yes