Elleason Joshua G.

Francisco
2E-Pharmacy

Recombinant DNA- a DNA in which

one or more segments or genes have
been inserted, either naturally or by
laboratory manipulation, from a
different molecule or from another
part of the same molecule, resulting in
a new genetic combination.
2. Gene transfer-the insertion of copies
of a gene into cells to induce
synthesis of the gene's product: the
desired gene may be microinjected
directly into the cell or it may be
inserted into the core of a virus by
gene splicing and the virus allowed to
infect the cell for replication of the
gene in the cell's DNA.
3. Restriction endonucleases-enzymes
that hydrolyze double-stranded DNA at
specific spots on opposite strands,
also known as Restriction enzymes
are enzymes that cut a DNA molecule
at a particular place. They are
essential tools for recombinant DNA
technology. The enzyme "scans" a
DNA molecule, looking for a particular
sequence, usually of four to six
nucleotides.
4. Sticky ends- Longer overhangs are
called cohesive ends or sticky ends.
They are most often created by
restriction endonucleases when they
cut DNA. Very often they cut the two
DNA strands four base pairs from each
other, creating a four-base 5' overhang
in one molecule and a complementary

5' overhang in the other, They are

short,single stranded stretches at the
ends of double stranded DNA: they
can provide sites to which DNA
molecules with sticky ends can be
linked
April 29, 2016

5. Blunt ends- a Non-blunt ends are

created by various overhangs
,overhang is a stretch of unpaired
nucleotides in the end of a DNA
molecule. These unpaired nucleotides
can be in either strand, creating either
3' or 5' overhangs. These overhangs
are in most cases palindromic.

6. Vectors- Carrier molecule for

transfer of genes in DNA
recombination, In molecular cloning, a
vector is a DNA molecule used as a
vehicle to artificially carry foreign
genetic material into another cell,
where it can be replicated and/or
expressed. A vector containing foreign
DNA is termed recombinant DNA. The
four major types of vectors are
plasmids, viral vectors, cosmids, and
artificial chromosomes. Of these, the
most commonly used vectors are
plasmids. Common to all engineered
vectors are an origin of replication, a
multicloning site, and a selectable
marker. It is generally a DNA sequence

that consists of an insert (transgene)

and a larger sequence that serves as
the "backbone" of the vector. The
purpose of a vector which it transfers
genetic information to another living
cell is a typically to isolate, multiply, or
express the insert in the target cell.
7. Genetic engineering- the process of
manipulating the genome of an
organism to achieve a desired end, is
the modification of an organism's
genetic composition by artificial
means, often involving the transfer of
specific traits, or genes, from one
organism into a plant or animal of an
entirely different species.
8. Gene cloning- the introduction of a
selection of DNA into a genome in
which it can be reproduced many
times, it is the process in which a gene
is copied (cloned) out of DNA
extracted from an organism. DNA is
extracted from an organism, all of its
genes are extracted at one time. This
DNA, which contains thousands of
different genes.
9. Pcr and steps and components of
pcr and instrument use and 5
applications of pcr

Bio Rad Pcr Machine

Components
Magnesium chloride: .5-2.5mM
Buffer: pH 8.3-8.8
dNTPs: 20-200uM
Primers : 0.1-0.5uM
DNA Polymerase : 1-2.5 units
Target DNA : < 1ug

Application of PCR
-Neisseria gonorrhoea
-Chlamydia trachomatis
-HIV-1
-Factor V Leiden
-Forensic testing and many others