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Q1. In the week two laboratory class, you set-up three polymerase chain rea...
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In the week two
laboratory class, you set-up three polymerase chain reaction (PCR) reactions. Complete
the steps below (A-D) to upload and interpret your PCR results.
A) PCR RESULTS UPLOAD: Upload the agarose gel image of your PCR results to the file upload box. Record which gel lanes correspond to your experimental reaction, positive control reaction and negative control reaction in the provided table.
B) NEGATIVE CONTROL REACTION
The expected outcome for the negative control reaction is no visible DNA bands.
i) Did you observe any visible bands in your negative control? (YES/NO) (1 mark)
If NO, proceed to the next question (ii). If YES, explain whether or not you can have confidence in the results obtained for the positive control reaction and experimental reaction. In your answer, consider what the size and appearance of the observed band(s) may indicate.
C) POSITIVE CONTROL REACTION
The expected outcome for the positive control reaction is a single DNA band at 187 bp.
ii) Did your positive control produce a band of the expected size? (YES/NO) (1 mark)
If YES, proceed to the next question (iii). If NO, suggest one or more sources of error or procedural issues that could have resulted in failed amplification of the target sequence.
iii) Did your positive control produce an additional band(s) at unexpected sizes? (YES/NO) (1 mark)
If NO, proceed to the next question (iv). If YES, explain whether or not these bands represent non-specific amplification. If they do represent non-specific amplification, suggest one or more adjustments to the reaction mix or thermal cycling conditions that could improve the specificity of the PCR.
D) EXPERIMENTAL REACTION
The expected outcome for the experimental reaction is a single DNA band at 187 bp.
iv) Did your experimental reaction produce a band of the expected size? (YES/NO) (1 mark)
If YES, proceed to the next question (v). If NO, suggest one or more sources of error or procedural issues that could have resulted in failed amplification of the target sequence.
v ) Did your experimental reaction produce an additional band(s) at unexpected sizes? (YES/NO) (1 mark)
If NO, proceed to the next quiz question. If YES, explain whether or not these bands represent non-specific amplification. If they do represent non-specific amplification, suggest one or more adjustments to the reaction mix or thermal cycling conditions that could improve the specificity of the PCR.