NEED HELP WITH THESE QUESTIONS. PLEASE ANSWER ALL AND EXPLAIN AS WELL. THANKSSSS
ID: 267012 • Letter: N
Question
NEED HELP WITH THESE QUESTIONS. PLEASE ANSWER ALL AND EXPLAIN AS WELL. THANKSSSSSSS
1. You want to clone a gene from a donor vector to a host vector. List the correct order of events in the process of cloning a. Perform ligation reaction of cloned gene and host vector. b. Perform double digestion of both donor and host vectors with the 2 restriction enzymes c. Examine donor and host vectors for restriction sites d. Purify cloned gene from donor vector and purify host vector 2. Please list True (T) or False (F) for each question Sanger sequencing involves RNA synthesis of a complementary strand that to the target of interest Sanger sequencing involves addition of dideoxy ATP (ddATP), dideoxy CTP (ddCTP) dideoxy GTP (ddGTP), dideoxy TTP (ddTTP) to the polymerase reactionn ddNTP molecules each inhibit polymerase activity which results in chain termination Fragments of varying length and sequence are analyzed with electrophoresis and autoradiography Comparison of each oligomer on the gel can lead to obtaining of the sequence one nucleotide at a time a. b. c. d. e. 3. Please list True (T) or False (F) for each question In site directed mutagenesis, a mismatch between the cloned gene and an oligonucleotide primer is created The oligonucleotide is not annealed during polymerase activity A closed circular duplex is created using DNA polymerase The mutant gene is generated when the closed circular duplex is melted, and new complementary mutant gene is synthesized a. b. c. d. 4. Please list True (T) or False (F) for each question PCR is a highly sensitive technique for amplifying PCR PCR results in linear amplification rather than exponential synthesis of DNA The requirements for the PCR reaction are template, primer, dNTPs, and polymerase A DNA polymerase that is derived from thermus aquaticus is used so that DNA polymerase can be used at high temperatures PCR can be used for forensics, amplification of specific DNA sequences, and qRT-PCR a. b. c. d. e.Explanation / Answer
2. Sanger sequencing involves RNA synthesis of a complementary strand that to the target of interest False: Sanger sequencing is the process of determining the sequence of nucleotides (As, Ts, Cs, and Gs) in a piece of DNA.
sanger sequencing involves addition of dideoxy ATP (ddATP), dideoxy CTP (ddCTP), dideoxy GTP (ddGTP), dideoxy TTP (ddTTP) to the polymerase reaction
TRUE,
ddNTP molecules each inhibit polymerase activity which results in chain termination
TRUE: Once a dideoxy nucleotide ddNTP has been added to the chain, there is no hydroxyl available and no further nucleotides can be added. The chain ends with the dideoxy nucleotide, which is marked with a particular color of dye depending on the base (A, T, C or G) that it carries.
fragments of varying length and sequences are analysed with electrophoresis and autoradiography
TRUE
comparision of each oligomer on the gel can lead to obtaining of the sequence one nucleotide at a time
TRUE
3. In site directed mutagenesis, a mismatch between the cloned gene and an oligonucleotide primer is created TRUE
oiligonucleotide is not annealed during polymerase activity FALSE
A closed circular duplex is created using DNA polymerase TRUE
The mutant gene is generated when the closed circular duplex is melted and new complementary mutant gene is synthesized.TRUE
PCR is a highly sensitive technique for amplifying PCR TRUE
PCR results in linear amplification rather than exponential synthesis of DNA FALSE
The requirement of PCR reaction are template, primer,dNTP and polymerase TRUE
A DNA polymerase that is derived from thermos aquiticus is used so that DNA polymerase can be used at high temperatures TRUE. it is stable at high temperature
PCR can be used for forensics, amplification of specific DNA sequences and qRT-PCR
TRUE PCR is useful in forensic to ensure the enough quantity of DNA
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