The purpose of the lysis procedure is:

to sequence the 16S rRNA gene

to separate the pieces of nucleic acid from one another

to break open the bacterial cells to release the geneticmaterial

to amplify the 16S rRNA gene

Match the components of HotShot buffer with theirrole.

NaOH    

           removes anions so they cannot be used as cofactors by enzymes,increases pH to denature nucleases, disrupts lipid membranes,removes cations so they cannot be used as cofactors by enzymes, ordecreases pH to denature nucleases

Detergent

           removes anions so they cannot be used as cofactors by enzymes,increases pH to denature nucleases, disrupts lipid membranes,removes cations so they cannot be used as cofactors by enzymes, ordecreases pH to denature nucleases

Chelator

            (removes cations so they cannot be used as cofactors byenzymes)

When taking some of your bacterial isolate to place inthe HotShot buffer you should take as much as can fit on the end ofthe pipette tip.

True OR False

The role of neutralization buffer is to:

restore the pH to approximately neutral

increase the pH to 10

introduce nucleases into the reaction

decrease the pH to 3

Nuclease-free water is added to the reaction mix to_____________________. Choose all that apply.

increase the volume of the solution without introducingnucleases which could degrade the nucleic acids

dilute the lysate so it is not too concentrated for laterprocedures

to stop the chemical reactions occuring in the solution

add nucleases to the reaction mix