Student_summary_gel_procedure

Student_summary_gel_procedure - negative control 3 What...

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Summary gel electrophoresis for DNA analysis lab #10 1 2 3 4 Sample Sample *Sample + DNase @ Sample not + DNase (contaminated incubated incubated with RNase) - DNase DNA ------ ----- band RNA ------ ------ 2 bands RNA ------ ------ TRNA Smear at lower end of gel (large diffuse band) _____________________________________________________________________________________ 1. Sample in lane 1 that is not incubated at 37 C shows complete DNA and RNA bands Expected result with Dnase is shown in lane 2.. They may see one to three RNA bands (two rRNAs middle of gel and tRNA in a smear at the bottom). 2. * What if there is RNase contamination in Dnase, lane 3 shows a possible result?
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Unformatted text preview: negative control. 3. @ What might your E. coli DNA preparation be contaminated with? Shows DNA prep is contaminated with Rnase in lane 4. Note that E. coli contains several enzymes that could digest DNA and/or RNA. Reactions 1 2 3 4 prep 1 prep 2 prep 1 prep1 + DNase- DNase H 2 O 8 µl 8 µl 5 µl 8 µl Nucleic Acid 10 µl 10 µl 10 µl 10 µl Buffer (10X) 2 µl 2 µl 2 µl 2 µl DNase--3 µl-20 20 20 20 5 6 7 8 prep 2 prep 2 tRNA tRNA + DNase + DNase + DNase-DNase Heated H 2 O 5 µl 5 µl 5 µl 8 µl Nucleic Acid 10 µl 10 µl 10 µl 10 µl Buffer (10X) 2 µl 2 µl 2 µl 2 µl DNase 3 µl 3 3 µl-20 20 20 20...
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