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1. Observe the photograph of the gel containing your sample. Orient the photograph with wells at the top. Interpret each lane of the gel. a. Scan across the photograph of your gel to get an impression of what you see in each lane. You should notice that virtually all experiment lanes contain one or two prominent bands. b. Now locate the lane containing the pBR322/BstNI marker on the left hand side of the gel. Working from the well, locate the bands corresponding to each restriction fragment: 1,857 bp, 1,058 bp, 929 bp, 383 bp, and 121 bp (may be faint). Alternatively, locate the lane containing the 100-bp ladder, with the fastest migrating band equal to 100 bp and each successive band 100 bp larger (100, 200, 300,400, etc.). c. The amplification products of the 35S promoter (162 bp) and of the tubulin gene (187 bp) should align between the 121-bp and 383-bp fragments of the pBR322/BstNI marker (or between the 100-bp and 200-bp markers of the 100-bp ladder). d. It is common to see one or two diffuse (fuzzy) bands of RNA and/or primer dimer at the bottom of the gel. RNA may be found at approximately the position of the 121-bp fragment of the pBR322/BstNI marker (or the 100-bp marker of the 100-bp ladder). RNA is the largest component of nucleic acid isolated from plant tissue, so some RNA may escape digestion by the RNase incorporated into the TE buffer in Part II of the experiment. Primer dimer is an artifact of the PCR reaction that results from two primers overlapping one another and amplifying themselves. Primer dimer is approximately 50 bp and should be in a position ahead of the 121-bp marker fragment. e. Additional faint bands, at other positions on the gel, occur when the primers bind to chromosomal loci other than 35S or tubulin, giving rise to "nonspecific" amplification products.

1. Observe the photograph of the gel containing your sample. Orient the photograph with wells at the top. Interpret each lane of the gel. a. Scan across the photograph of your gel to get an impression of what you see in each lane. You should notice that virtually all experiment lanes contain one or two prominent bands. b. Now locate the lane containing the pBR322/BstNI marker on the left hand side of the gel. Working from the well, locate the bands corresponding to each restriction fragment: 1,857 bp, 1,058 bp, 929 bp, 383 bp, and 121 bp (may be faint). Alternatively, locate the lane containing the 100-bp ladder, with the fastest migrating band equal to 100 bp and each successive band 100 bp larger (100, 200, 300,400, etc.). c. The amplification products of the 35S promoter (162 bp) and of the tubulin gene (187 bp) should align between the 121-bp and 383-bp fragments of the pBR322/BstNI marker (or between the 100-bp and 200-bp markers of the 100-bp ladder). d. It is common to see one or two diffuse (fuzzy) bands of RNA and/or primer dimer at the bottom of the gel. RNA may be found at approximately the position of the 121-bp fragment of the pBR322/BstNI marker (or the 100-bp marker of the 100-bp ladder). RNA is the largest component of nucleic acid isolated from plant tissue, so some RNA may escape digestion by the RNase incorporated into the TE buffer in Part II of the experiment. Primer dimer is an artifact of the PCR reaction that results from two primers overlapping one another and amplifying themselves. Primer dimer is approximately 50 bp and should be in a position ahead of the 121-bp marker fragment. e. Additional faint bands, at other positions on the gel, occur when the primers bind to chromosomal loci other than 35S or tubulin, giving rise to "nonspecific" amplification products.

Human Anatomy & Physiology (11th Edition)
Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
Section: Chapter Questions
Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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1. Observe the photograph of the gel containing your sample. Orient the photograph with wells at the top. Interpret each lane of the gel. a. Scan across the photograph of your gel to get an impression of what you see in each lane. You should notice that virtually all experiment lanes contain one or two prominent bands. b. Now locate the lane containing the PBR322/BSTNI marker on the left hand side of the gel. Working from the well, locate the bands corresponding to each restriction fragment: 1,857 bp, 1,058 bp, 929 bp, 383 bp, and 121 bp (may be faint). Alternatively, locate the lane containing the 100-bp ladder, with the fastest migrating band equal to 100 bp and each successive band 100 bp larger (100, 200, 300 ,400, etc.). c. The amplification products of the 35S promoter (162 bp) and of the tubulin gene (187 bp) should align between the 121-bp and 383-bp fragments of the PBR322/BstNI marker (or between the 100-bp and 200-bp markers of the 100-bp ladder). d. It is common to see one or two diffuse (fuzzy) bands of RNA and/or primer dimer at the bottom of the gel. RNA may be found at approximately the position of the 121-bp fragment of the pBR322/BSTNI marker (or the 100-bp marker of the 100-bp ladder). RNA is the largest component of nucleic acid isolated from plant tissue, so some RNA may escape digestion by the RNase incorporated into the TE buffer in Part II of the experiment. Primer dimer is an artifact of the PCR reaction that results from two primers overlapping one another and amplifying themselves. Primer dimer is approximately 50 bp and should be in a position ahead of the 121-bp marker fragment. e. Additional faint bands, at other positions on the gel, occur when the primers bind to chromosomal loci other than 35S or tubulin, giving rise to "nonspecific" amplification products.
Transcribed Image Text:1. Observe the photograph of the gel containing your sample. Orient the photograph with wells at the top. Interpret each lane of the gel. a. Scan across the photograph of your gel to get an impression of what you see in each lane. You should notice that virtually all experiment lanes contain one or two prominent bands. b. Now locate the lane containing the PBR322/BSTNI marker on the left hand side of the gel. Working from the well, locate the bands corresponding to each restriction fragment: 1,857 bp, 1,058 bp, 929 bp, 383 bp, and 121 bp (may be faint). Alternatively, locate the lane containing the 100-bp ladder, with the fastest migrating band equal to 100 bp and each successive band 100 bp larger (100, 200, 300 ,400, etc.). c. The amplification products of the 35S promoter (162 bp) and of the tubulin gene (187 bp) should align between the 121-bp and 383-bp fragments of the PBR322/BstNI marker (or between the 100-bp and 200-bp markers of the 100-bp ladder). d. It is common to see one or two diffuse (fuzzy) bands of RNA and/or primer dimer at the bottom of the gel. RNA may be found at approximately the position of the 121-bp fragment of the pBR322/BSTNI marker (or the 100-bp marker of the 100-bp ladder). RNA is the largest component of nucleic acid isolated from plant tissue, so some RNA may escape digestion by the RNase incorporated into the TE buffer in Part II of the experiment. Primer dimer is an artifact of the PCR reaction that results from two primers overlapping one another and amplifying themselves. Primer dimer is approximately 50 bp and should be in a position ahead of the 121-bp marker fragment. e. Additional faint bands, at other positions on the gel, occur when the primers bind to chromosomal loci other than 35S or tubulin, giving rise to "nonspecific" amplification products.
BD II
Transcribed Image Text:BD II
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