Lab 3

under what conditions will the E Coli K- 12 glow green? (Recombinant DNA Technology lab)

+ Arabinose

What is the purpose of a buffer? (DNA Lab)

-To provide ions for electrophoresis. - keeps ph constant . -contains stain (bromphenol blue) to determine length of electrophoresis -contains ficoll to make sample more dense.

PCR 3 steps (DNA Lab)

1) denature DNA 95 degrees 2) anneal primer 3)polymerization

what is a pGlo? What abilities does pGlo transfer to bacteria? How ? (Recombinant DNA Technology lab)

1)The pGLO plasmid is an engineered plasmid used in biotechnology as a vector for creating genetically modified organisms. light turns green because the gene they have. . 2) Because it is ampicillin resistant Florescent is transferred. 3)The bacteria's increased, antibiotic resistant, it has survival abilities.

What is a probe?

A labeled nucleic used to identify a complementary region in a clone or genome

What is a plasmid and its function?

A plasmid is a structure that physically separated from your chromosomes DNA. function- Plasmid can replicate independently

What is a promoter?

A promoter is a regulatory region located at the beginning of a gene

Why would you expect to find cell growth on the Lura nutrient agar plate? (LB plate) (Recombinant DNA Technology lab)

Active bacteria without amp. to stop growth. bacteria is alive.

a restriction enzyme is a

Bacterial enzyme that cuts DNA at specific sites.

How does gel electrophoresis work? What is accomplished?

Because each DNA molecule is negatively charged, it can be pulled through the gel by an electric field. Small DNA molecules move more quickly through the gel than larger DNA molecules

How do normal cells become tumor cells? (Carcinogenesis lab )

Because they mutate

What is the purpose of the CaCl2 step in the lab? (DNA Lab)

Calcium chloride transformation is a laboratory technique in prokaryotic bacterial cell biology. It increase the ability of a prokaryotic cell to incorporate plasmid DNA allowing them to be genetically transformed.

In recombinant DNA techniques, primers and DNA fragments are linked together by

DNA ligase

In recombinant DNA techniques, the gaps in the paired DNA fragments are closed by?

DNA ligase

DNA ligase and its function?

DNA ligase is linking enzyme essential for DNA replication, catalyzes and completes the DNA backbone by forming covalent phosphodiester

what is DNA polymerase and its function?

DNA polymerase is an enzyme that catalyzes the synthesis of dna using a template dna strand an nucleotides

Vectors can transfer

DNA sequences into bacterial host cells for cloning.

What do restriction enzymes contribute to the forensic process? How do they work? (DNA Lab)

Eco RI cuts the DNA sequence between the sugar and the phosphate in the sugar and the phosphate in the sugar- phosphate backbone of DNA.

(Recombinant DNA Technology lab) How do the experimental plates demonstrates that transformation has occurred?

Growing colonies under UV light.

How does Heat shock affect plasmid transformation? (Recombinant DNA Technology lab)

Heat shock makes bacteria to take in plasmid.

What are the dependent and independent variables? (Recombinant DNA Technology)

Independent is plasmid Dependent is glow

(Carcinogenesis) What are the dependent and independent variables?

Independent variable carcinogenesis is bacterium Dependent variable is the tumor formation

What is the purpose of restriction enzymes and how do they work?

It recognizes a short and specific nucleotide sequence that cuts the DNA only at the specific sites called a target sequence

What is restriction enzyme? How were restriction enzymes used to construct pGlo? (Recombinant DNA Technology lab)

Molecular scissors, sticky ends. carrying the genetic code of the organism, DNA cells pick up DNA from environment. Arac black RNA from being transferred and arabonase breaks arc down.

(Carcinogenesis lab ) What are mutagens?

Mutagens are physical and chemical agents that cause mutations

Did you expect to find cell growth on the ampicillin Luria plate? (Recombinant DNA Technology lab)

No because AMP restrict the growth of the bacteria without plasmid.

what is RNA polymerase and its functions

RNA polymerase is an enzyme that is responsible for making RNA from a DNA template. RNA polymerase functions is to produce a complementary RNA strand

Explain DNA fingerprinting

Repetitive regions in non-coding DNA, also called "junk" DNA are used for DNA fingerprinting.

RFLP (DNA Lab)

Restriction Fragment Length polymorphism

What is the role of a tumor suppressor, oncogene or DNA repair gene in the development of cancerous tissue? (Carcinogenesis lab )

The role of a tumor suppressor genes are normal gene that slow down cell division, but oncogene or DNA repair DNA gene tells cells when to die or a process known as apoptosis (programmed cell death)

A. tumfaciens may contain a small plasmid called? (Carcinogenesis lab)

Ti- Plasmid (tumor- inducing plasmid) which can transfer to damaged plant tissue and cause disorganized cellar growth (neoplasia) in the plant cell.

what agent caused the cancerous growths observed?

Ti- plasmid caused cancerous growth (agar bacterium)

How was the carcinogen delivered to the carrot tissue?

a cambian

Most often, genomic library screenings uses

a labeled, complementary single stranded RNA or DNA molecule called a probe.

Dysplasia? (Carcinogenesis lab)

abnormal maturation of cells in a tissue. Cell maturation and differentiation are delayed. (no longer makes Carotene)

independent variable (carrot lab)

agar bacterium

plasmid transformation

altering the genetic code of an organism,and change its survival abilities

VNTS's and STR's? (DNA Lab)

are repetitive sequences of different lengths.

In bacterial genetics, what is meant by the term Transformation? (Recombinant DNA Technology lab)

carrying the genetic code of the organism. DNA cells pick up DNA from environment.

When Ti genes are integrated, the plant cell looses its ability to differentiate properly_________and to control cellular growth?

dysplasia

During the process of gel electrophoresis, the DNA fragments migrate due to the effects of

electrical charge.

polymerization via nucleotides

elongation

plasmids are short sequences od DNA used as primers in PCR? True or false?

false

true or false? all tumors are cancerous?

false- a tumor is only cancerous only if it infiltrates nearby tissue

true or false? cancer begins when a cell divides less frequently than it should

false- cancer behind when a cell divides more often than it should

T or F? plasmids are short sequences of DNA used as primers in PCR?

fasle

Plasmids are used by Genetic Engineers to insert human gene codes into bacterial cells. Describe some benefits humanity can hope to gain by such an insertion.? (Recombinant DNA Technology lab)

genetically modified organism.

Neoplasia? (Carcinogenesis lab)

greek for "new growth". Abnormal proliferation of cells could be benign or malignant

What is calcium chloride transformation? What does it accomplish? (Recombinant DNA Technology lab)

increase the ability to take up plasmid, neutralize membrane of bacteria.

Agrobacterium tumefaciens (also called Rhizobium radiobacter)? (Carcinogenesis lab)

is a bacterial strain, which is able to cause the development of tumors in plants.

What is caner? (Carcinogenesis lab)

is a disease of genes. Cancer cells are able to override important cell cycle control mechanism.

why is the cancer known as Crown Gall Tumor?

it stains differently

What is the purpose of the loading dye? (DNA Lab)

its more dense, contains dye to help determine length of electrophorus.

Is DNA positively charged or negatively charged? (DNA Lab)

its negatively charged and it moves towards + charge.

malignancy? (Carcinogenesis lab)

killing of surrounding tissue

Be able to analyze a DNA fingerprint gel.? (DNA Lab)

look at diagrams on page 63. in lab manual

if a cell begins to divide in an uncontrolled way, it

may form a benign tumor

metastasis? (Carcinogenesis lab)

metastasize spread of disease from one organ to another.

PCR (DNA Lab)

polymerase chain reaction- amplifying DNA

During PCR__________serve as a starting point for the synthesis of the new DNA strand.

primers

benign? (Carcinogenesis lab)

restricted to originating area

plasmids, which are found within microorganisms can be cut open by (Recombinant DNA Technology lab)

restriction enzymes

What is E. coli K-12? (Recombinant DNA Technology lab)

safety strain, it cannot grow in your intestine.

In the creation of a DNA profile_________are typically used

short tandem repeats

What are STRs and VNTRs?

short tandem repeats and variable number tandem repeats.

Receptive regions is non-coding (units) in "junk DNA"? (DNA Lab)

short tandem repeats.

carolina Blue- methylene blue

staining dye- methylein blue

Southern blot? (DNA Lab)

the DNA is visualized afterwards either using a stain, or radioactive probe in a southern blot.

Transformation (Carcinogenesis lab)

the introduction of external DNA into a cell

PCR enables what? (DNA Lab)

the investigation team to take a minute quantities of sample and increase the amount of DNA obtained from the sample.

dependent variable (carrot lab)

the tumor formation

Southern Blot is a method for

transferring DNA fragments from a gel to a membrane filter and used in hybridization experiments.

During gel electrophoresis, DNA fragments are separated by size. true or false?

true

Repeating the sequence of the polymerase chain reaction doubles the number of DNA fragments each time. true or false?

true

true or false? one cells may develop mutations in cancer causing agents

true

true or false? the process of a tumor spreading is called metastasis?

true

Ananplasia? (Carcinogenesis lab)

undifferentiated growth in a malignant neoplasm lack of differentiation, tumor cells invade neighboring

VNTR's? (DNA Lab)

variable number tandem repeats

experimental

with agar plate + bacterium