1. Sample buffer contains glycerol, xylene cyanol and bromophenol blue. It provides three important functions.
i. It provides density to the sample
ii. It helps in monitoring the gel running status
iii. If the sample volume is too low (Ex: 1uL or 0.5 uL), we can dilute it with the sample buffer so that it can be easily loaded into the gel lane.
2. If the DNA is not mixed with sample buffer,
i. The DNA would get diffused into the tank buffer/gel. SO, we can not observe/obtain crisp bands (The quantity also gets reduced as some sample diffuses out).
ii. We can not monitor the gel running status.
3. The blue dye runs at ~50 bp region. So, we can approximately guess the position of our interested DNA bragment on the gel using the blue dye as a control.
The xylene cyanol band (Light green/blusih green) runs approximately at 3kb.
1. What is the purpose of the sample buffer? 2. What would happen if you did...
Before you add your DNA sample to your well, what do you have to add to the sample? O primer O probe O loading buffer (aka loading dye) O size standard (aka marker DNA) Question 10 0.5 pts If you want to estimate the sizes of the DNA fragments in your gel, what do you have to load alongside your DNA samples? primer probe loading buffer (AKA loading dye) size standard (AKA marker DNA)
The following is the recipe for 4X sample loading buffer: 1. 4X stacking gel buffer-10 mL 2. 10% SDS- 18 mL 3. beta-mercaptoethanol- 2 mL 4. glycerol- 20 mL 5. Dash bromophenol blue (assume no volume change) Questions: 1. What is the final concentration of SDS in the solution of 4X sample loading buffer? 2. What is the purpose of the following reagents in 4X sample loading buffer: a) SDS? b) Beta-mercaptoethanol? c) glycerol? d) Bromophenol blue?
The following is the recipe for 4X sample loading buffer: 4X stacking gel buffer – 10ml 10% SDS – 18 ml Beta-mercaptoethanol – 2 ml Glycerol – 20ml Dash bromophenol blue (assume no volume change) A) What is the final concentration of SDS in the solution of 4X sample loading buffer? B) What is the purpose of the following reagents in 4X sample loading buffer: i) SDS? ii) Beta-mercaptoethanol? iii) Glycerol? iv) Bromophenol blue?
what would happen to the proteins in your gel if you did not add SDS-PAGE?
20- In cheek DNA extraction procedure, what is the purpose of heating a. To speed up the DNA precipitation b. To hydrolyze protein content c. To lysis cell d. To separate DNA fragments 21- What is the reason of using Taq DNA Polymerase in most of the PCR reactions. a. Taq polymerase is sensitive to high temperature b. Taq is very cheep to produce c. Taq can tolerate DNA denaturation temperature d. a, and b Assay questions 5 points each...
6. When performing agarose gel electrophoresis, how much 6X loading dye should you add to a 8 uL DNA sample before loading it onto the gel? (1 pt)
please! Assay questions 5 points each 1-What RFLP stand for and what RFLP analysis test for? 2-Why do you add sample loading buffer to DNA sample prior to loading sample into agarose gel. 3-In cheek DNA extraction procedure, name two steps that helped to break (lysis) the cheek cells. 4- Would a shorter DNA fragment move faster or slower through agarose gel, why.
nomal No Spac... Heading 1 Paragraph JULLL HDBl AaBbCcDAoBbced Heading 2 Title Subtitle Subtle Em Styles 1-What RFLP stand for and what RFLP analysis test for? 2-Why do you add sample loading buffer to DNA sample prior to loading sample into agarose gel. 3 In cheek DNA extraction procedure, name two steps that helped to break (lysis) the cheek cells. 4. Would a shorter DNA fragment move faster or slower through agarose gel, why.
What is the purpose in a PCR reaction for each of the following reagents? Taq Taq buffer dNTPs Forward primer Reverse primer Genomic DNA What do you think would happen if you forgot to add your Reverse primer when you did a PCR?
So far in class I have learned c1v1=c2v2....it is said v1 is usually unknown. I am unsure how to solve this problem. 1.You have 23 uL of plasmid DNA that you want to run on a gel, but you only have 7X tracking dye. How much 7X tracking dye would you add to your sample before loading it on the gel?