After transduction we move to mutagenesis analysis. Using the transduction plate，PCR I found that lacZ，fliE，nagC,ompA all show successful transduction and successful PCR. But for DNA sequencing some clonies are not correct. Expected size of PCR are lacZ 304，fliE 255，rpoA 420，nagC 329，ompA 731，lexA 350，kan cassette 549，tet cassette 265. Please indicate which mutation were successful and success why some were not from the size about. indicate at what stage any of the mutagenesis failed.

Which of the following is true for nested PCR?   1. In the second round of PCR, primers should  anneal  only to the target site   2. Two primers are used to ensure that one or the other will anneal.   3. Both primers anneal only to the target site, the second round is to get more DNA   4. In the first round of PCR, primers should only anneal to the target site

CRISPR One of the more recent advances in biotechnology is the development of the CRISPR gene modification tool. Match the following descriptions the the key players in this technology. NOTE: If you want to change your selection, you'll need to delete the one you already chose. After you delete it, the list of choices will pop back up and you can make a different choice. CRISPR technology Specialized stretches of DNA with nucleotide repeats and Cas9 protein spacers Enzyme that cuts DNA crRNA Guides the enzyme to the target site CRISPR DNA Adapted from the natural defense mechanism of bacteria and archaea

You buy a new PCR kit.  The starting concentration for each reagent is as follows: the primers are 200 mM (mmole/L), the PCR buffer + dNTPs is 150 mM (mmole/L), and the polymerase 15 units/mL.      In the protocol manual, it states that you want the final concentration (FC) of primers to be 40 mM each, the FC of PCR buffer + dNTPs to be 25 mM, and to have 4 units of the DNA polymerase in the final 50 mL PCR reaction.

Using the Figure below briefly describe how the amplification can be used in the cloning process of DNA.

Which of the following statements about PCR or RT-gPCR is correct? 1. Fluorescence is detected when the TagMan probe is cleaved by Taq polymerase (AMPLITAQ). 2. PCR uses the natural process of transcription to amplify DNA. 3. Both PCR and RT-qPCR require forward and reverse primers. 4. Fluorescence is detected when the reporter dye is in close proximity to the quencher dye. A. 1, 2 and 3 B. 1 and 3 C. 2 and 4 D. 4 only E. All of 1, 2, 3 and 4 are correct

In the following "gene library" cloning experiment Digested genomic DNA AmpR gene TCR gene TCR is tetracycline resistant marker, AmpR is ampicillin resistant marker and BamHI is the unique restriction enzyme on plasmid. A PhD student digests/cuts the plasmids with BamHI restriction enzyme and the genomic DNA with EcoRI restriction enzyme. After performing the cloning experiment and obtaining colonies on a selection plate, the obtained cells will be ..... (Hint: this question is even more challenging; the PhD student was later demoted to an MSc student). a) resistant to ampicillin and tetracycline b) sensitive to tetracycline and ampicillin c) resistant to tetracycline and sensitive to ampicillin d) resistant to ampicillin and sensitive to tetracycline e) sensitive to ampicillin and tetracycline BamHI

First option is : was not  or was Second option is : can or cannot Thrid option: cut, cut and amplified, amplifed