What is the main reason for using a cDNA library rather than a genomic library to isolate a human gene from which you wish to make large quantities of the human protein in bacteria? How will you identify clones of interest?

Your colleague wants to align the complete sequence of the collagen gene from humans to the homologous complete gene in rats. She can't decide if she should use the alignment program Needle or Water, and turns to you for help. Does it make a difference which program she uses, what advice would you give her?

You have assembled a new prokaryotic genome and are annotating the genes.  If you were to search for highly conserved sequences necessary for normal gene expression, what type of annotation are you doing? Group of answer choices ad hoc annotation homology based annotation preliminary annotation ab initio annotation

A possible forward primer to be designed is a primer that binds to the region of the OXA-M290 gene where the start codon is located, and the reverse primer is the primer that binds to the region of the gene where the stop codon is located.  In order to clone the PCR product generated using these primers into pET-28a, extra nucleotides must be added to the 5' ends of the primers. These extra nucleotides will contain the recognition sites for the restriction enzymes that are used for cloning. When the DNA produced by PCR using these primers is treated with the restriction enzymes, this will generate sticky ends at the ends of the PCR product. If pET-28a is digested with the same restriction enzymes, the sticky ends of the PCR product will bind to the sticky ends on the digested pET-28a, allowing the two DNA molecules to be connected together by DNA ligase. Which primer (forward or reverse) should contain the SacI recognition site, and which primer (forward or reverse) should contain the…

Now you have the gene sequence. Now you would like to clone it into an expression vector to grow up in a bacterial system. Because you're going to use bacteria to generate protein from a eukaryote, the mammoth, you need to get rid of introns from your sequence. How do you do that?     Bioinformatically, I look for splice-site sequences and branch-point adenines and predict intron-exon boundaries     I use a comparative genomic approach and use sequence homology with the genome of a closely related species     I use a comparative genomic approach and use sequence homology with the genome of a distantly related species     Both A and B     Both B and C   Why did you bother to identify the introns?     So that I could include them in the sequence to understand intron function.     So that I could exclude them from the sequence because prokaryotes don't have spliceosomal machinery.     So that I could see how introns affect protein folding.

You are engineering a new vector that contains a screenable marker that can be used for blue/white screening of successful clones. For each site (1, 2, and 3) on the cloning vector below, describe why it would or would not be a good place for you to put the polylinker to facilitate blue/white screening. You can assume that the polylinker itself will not interfere with coding sequence in that region. In other words, the polylinker length will be a multiple of 3 nucleotides, will not contain a stop codon, and any amino acids translated will not affect the activity of the protein in that region. The arrows indicate the direction of transcription for the gene.

What is the major difference between the strategies of map based sequencing and shotgun sequencing?

This is from the Wilson et al. article.

Use this information to match the list of probes to the 3 samples below: You have isolated several DNA sequences from a variety of mouse tissues. You have labeled each one of them with a radioisotope and will use them as probes on blots of several DNA and RNA somples. Below are a list of all the probes you generated (probes A through E) and a list of all the DNA and RNA samples that you will analyze (Samples 1 through 3) Beside each sample, write the letters corresponding to oll the probes that will bind to a complementary sequence in that sample. These responses are graded all or nothing! List of probes Probe A: promoter sequence of a gene that is only expressed in the nervous system Probe B: promoter sequence of one of the genes encoding a ubiquitously expressed histone protein Probe C: coding sequence of a gene that is only expressed in the nervous system Probe D: coding sequence of one of the genes encoding a ubiquitously expressed histone protein Probe E: intron of a gene that is…

If you were to sequence a human genome today, how would the sequencing differ from that done during the Human Genome Project? Choose only the best answer. You would still use Sanger sequencing. You would still assemble the sequencing reads into a genome using bioinformatics. You still need to clone the DNA fragments prior to sequencing them. All three of the other statements are true.