What are the primary components of a typical PCR reaction, and what does each of these components do?

BIOL3251 GENETICS LAB FALL 2018

ASSIGNMENT 4 Name:__________________________ PCR: Polymerase Chain Reaction Lab section:_____________________

The polymerase chain reaction, or PCR, allows us to make many copies of targeted regions of the genome for a number of different downstream applications. In this project, you are PCR-amplifying your mitochondrial HVR1 region so that you can obtain the DNA sequence for that region. The ultimate goal is to determine your haplotype or haplogroup based on that region, which will provide you with insight into your deep ancestry.

Consider the class discussions you had on PCR this week. Use this information to answer the following questions:

1. What are the primary components of a typical PCR reaction, and what does each of these components do?
 
 
 
 
 
 
 
 
 


2. How can these components be adjusted to optimize, or troubleshoot, a reaction that may not be yielding positive results?
 
 
 
 
 
 
 
 
 


3. What are the main steps of a thermal cycling profile, and how can these steps be adjusted to optimize, or troubleshoot, a reaction that may not be yielding positive results?
 
 
 


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BIOL3251 GENETICS LAB FALL 2018

The entire human mitochondrial genome is just over 16,500 base pairs (bp) in length, and it is organized as a single, circular chromosome. The Cambridge Reference Sequence, fully annotated and downloaded from GenBank, is provided below. You will use it to complete the follow questions.

4. Using a highlighter, colored pen, or colored pencil, mark and label the entire HVR 1 and HVR 2 regions on the Cambridge Reference Sequence below.
 


5. On the CRS below, mark and label the priming sites for the left and right primers used in today’s PCR experiment. Use a different color than was used for marking the entire HVR1 and HVR2 regions.
 
 Primer name Primer sequence

mtDNA_PCR_L 5’-tcattggacaagtagcatcc-3’ mtDNA_PCR_R 5’-aggctaagcgttttgagctg-3’
 
 Hints:

1. The primers are designed to amplify HVR1 and HVR2 as one contiguous unit.

2. The right primer is the reverse complement of its intended priming site. 3. The primers were designed to anneal (or bind) just outside of the HVR

regions to ensure that the entire region would be amplified and sequenced. 4. The primers were designed by Dr. Cahoon specifically for this class project,

so Google searching is pointless (just saving you the time and frustration :). 


6. What size amplicon (i.e., PCR product) do you expect these primers to generate?

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BIOL3251 GENETICS LAB FALL 2018

Homo sapiens mitochondrion, complete genome NCBI Reference Sequence: NC_012920.1 – full details of the sequence submission with all annotations can be obtained from http://www.ncbi.nlm.nih.gov/nuccore/NC_012920.

Human Mitochondrial Sequence and Annotation aka the Cambridge Reference Sequence (CRS)

This sequence was derived from DNA isolated from placental tissue collected from a caucasian living in Great Britain and was first published in 1981.

The mitochondrial haplotype given to this sequence is H1c

ANNOTATION:

Region or Gene Location Gene product HVR2 1 – 569 hypervariable region 2 TRNF 577..647 tRNA phenylalanine RNR1 648..1601 12S ribosomal RNA TRNV 1602..1670 tRNA valine RNR2 1671..3229 mitochondrially encoded 16S RNA TRNL1 3230..3304 tRNA leucine 1 ND1 3307..4262 NADH dehydrogenase 1 TRNI 4263..4331 tRNA isoleucine TRNQ 4329..4400 tRNA glutamine TRNM 4402..4469 tRNA methionine ND2 4470..5511 NADH dehydrogenase 2 TRNW 5512..5579 tRNA tryptophan TRNA 5587..5655 tRNA alanine TRNN 5657..5729 tRNA asparagine TRNC 5761..5826 tRNA cysteine TRNY 5826..5891 tRNA tyrosine COX1 5904..7445 cytochrome c oxidase I TRNS1 7446..7514 tRNA serine 1 TRND 7518..7585 tRNA aspartic acid COX2 7586..8269 cytochrome c oxidase II TRNK 8295..8364 tRNA lysine ATP8 8366..8572 ATP synthase 8 ATP6 8527..9207 ATP synthase 6 COX3 9207..9990 cytochrome c oxidase III TRNG 9991..10058 tRNA glycine ND3 10059..10404 NADH dehydrogenase 3 TRNR 10405..10469 tRNA arginine ND4L 10470..10766 NADH 4L ND4 10760..12137 NADH dehydrogenase 4 TRNH 12138..12206 tRNA histidine TRNS2 12207..12265 tRNA serine 2 TRNL 12266..12336 tRNA leucine 2 ND5 12337..14148 NADH dehydrogenase 5 ND6 14149..14673 NADH dehydrogenase 6 TRNE 14674..14742 tRNA glutamic acid CYTB 14747..15887 cytochrome b TRNT 15888..15953 tRNA threonine TRNP 15956..16023 tRNA proline HVR1 16001 – 16569 hypervariable region 1

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BIOL3251 GENETICS LAB FALL 2018

HUMAN mtDNA CAMBRIDGE REFERENCE SEQUENCE

1 gatcacaggt ctatcaccct attaaccact cacgggagct ctccatgcat ttggtatttt 61 cgtctggggg gtatgcacgc gatagcattg cgagacgctg gagccggagc accctatgtc 121 gcagtatctg tctttgattc ctgcctcatc ctattattta tcgcacctac gttcaatatt 181 acaggcgaac atacttacta aagtgtgtta attaattaat gcttgtagga cataataata 241 acaattgaat gtctgcacag ccactttcca cacagacatc ataacaaaaa atttccacca 301 aaccccccct cccccgcttc tggccacagc acttaaacac atctctgcca aaccccaaaa 361 acaaagaacc ctaacaccag cctaaccaga tttcaaattt tatcttttgg cggtatgcac 421 ttttaacagt caccccccaa ctaacacatt attttcccct cccactccca tactactaat 481 ctcatcaata caacccccgc ccatcctacc cagcacacac acaccgctgc taaccccata 541 ccccgaacca accaaacccc aaagacaccc cccacagttt atgtagctta cctcctcaaa 601 gcaatacact gaaaatgttt agacgggctc acatcacccc ataaacaaat aggtttggtc 661 ctagcctttc tattagctct tagtaagatt acacatgcaa gcatccccgt tccagtgagt 721 tcaccctcta aatcaccacg atcaaaagga acaagcatca agcacgcagc aatgcagctc 781 aaaacgctta gcctagccac acccccacgg gaaacagcag tgattaacct ttagcaataa 841 acgaaagttt aactaagcta tactaacccc agggttggtc aatttcgtgc cagccaccgc 901 ggtcacacga ttaacccaag tcaatagaag ccggcgtaaa gagtgtttta gatcaccccc 961 tccccaataa agctaaaact cacctgagtt gtaaaaaact ccagttgaca caaaatagac 1021 tacgaaagtg gctttaacat atctgaacac acaatagcta agacccaaac tgggattaga 1081 taccccacta tgcttagccc taaacctcaa cagttaaatc aacaaaactg ctcgccagaa 1141 cactacgagc cacagcttaa aactcaaagg acctggcggt gcttcatatc cctctagagg 1201 agcctgttct gtaatcgata aaccccgatc aacctcacca cctcttgctc agcctatata 1261 ccgccatctt cagcaaaccc tgatgaaggc tacaaagtaa gcgcaagtac ccacgtaaag 1321 acgttaggtc aaggtgtagc ccatgaggtg gcaagaaatg ggctacattt tctaccccag 1381 aaaactacga tagcccttat gaaacttaag ggtcgaaggt ggatttagca gtaaactaag 1441 agtagagtgc ttagttgaac agggccctga agcgcgtaca caccgcccgt caccctcctc 1501 aagtatactt caaaggacat ttaactaaaa cccctacgca tttatataga ggagacaagt 1561 cgtaacatgg taagtgtact ggaaagtgca cttggacgaa ccagagtgta gcttaacaca 1621 aagcacccaa cttacactta ggagatttca acttaacttg accgctctga gctaaaccta 1681 gccccaaacc cactccacct tactaccaga caaccttagc caaaccattt acccaaataa 1741 agtataggcg atagaaattg aaacctggcg caatagatat agtaccgcaa gggaaagatg 1801 aaaaattata accaagcata atatagcaag gactaacccc tataccttct gcataatgaa 1861 ttaactagaa ataactttgc aaggagagcc aaagctaaga cccccgaaac cagacgagct 1921 acctaagaac agctaaaaga gcacacccgt ctatgtagca aaatagtggg aagatttata 1981 ggtagaggcg acaaacctac cgagcctggt gatagctggt tgtccaagat agaatcttag 2041 ttcaacttta aatttgccca cagaaccctc taaatcccct tgtaaattta actgttagtc 2101 caaagaggaa cagctctttg gacactagga aaaaaccttg tagagagagt aaaaaattta 2161 acacccatag taggcctaaa agcagccacc aattaagaaa gcgttcaagc tcaacaccca 2221 ctacctaaaa aatcccaaac atataactga actcctcaca cccaattgga ccaatctatc 2281 accctataga agaactaatg ttagtataag taacatgaaa acattctcct ccgcataagc 2341 ctgcgtcaga ttaaaacact gaactgacaa ttaacagccc aatatctaca atcaaccaac 2401 aagtcattat taccctcact gtcaacccaa cacaggcatg ctcataagga aaggttaaaa 2461 aaagtaaaag gaactcggca aatcttaccc cgcctgttta ccaaaaacat cacctctagc 2521 atcaccagta ttagaggcac cgcctgccca gtgacacatg tttaacggcc gcggtaccct 2581 aaccgtgcaa aggtagcata atcacttgtt ccttaaatag ggacctgtat gaatggctcc 2641 acgagggttc agctgtctct tacttttaac cagtgaaatt gacctgcccg tgaagaggcg 2701 ggcataacac agcaagacga gaagacccta tggagcttta atttattaat gcaaacagta 2761 cctaacaaac ccacaggtcc taaactacca aacctgcatt aaaaatttcg gttggggcga 2821 cctcggagca gaacccaacc tccgagcagt acatgctaag acttcaccag tcaaagcgaa 2881 ctactatact caattgatcc aataacttga ccaacggaac aagttaccct agggataaca 2941 gcgcaatcct attctagagt ccatatcaac aatagggttt acgacctcga tgttggatca 3001 ggacatcccg atggtgcagc cgctattaaa ggttcgtttg ttcaacgatt aaagtcctac 3061 gtgatctgag ttcagaccgg agtaatccag gtcggtttct atctacnttc aaattcctcc 3121 ctgtacgaaa ggacaagaga aataaggcct acttcacaaa gcgccttccc ccgtaaatga 3181 tatcatctca acttagtatt atacccacac ccacccaaga acagggtttg ttaagatggc 3241 agagcccggt aatcgcataa aacttaaaac tttacagtca gaggttcaat tcctcttctt 3301 aacaacatac ccatggccaa cctcctactc ctcattgtac ccattctaat cgcaatggca 3361 ttcctaatgc ttaccgaacg aaaaattcta ggctatatac aactacgcaa aggccccaac 3421 gttgtaggcc cctacgggct actacaaccc ttcgctgacg ccataaaact cttcaccaaa 3481 gagcccctaa aacccgccac atctaccatc accctctaca tcaccgcccc gaccttagct 3541 ctcaccatcg ctcttctact atgaaccccc ctccccatac ccaaccccct ggtcaacctc 3601 aacctaggcc tcctatttat tctagccacc tctagcctag ccgtttactc aatcctctga

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BIOL3251 GENETICS LAB FALL 2018

3661 tcagggtgag catcaaactc aaactacgcc ctgatcggcg cactgcgagc agtagcccaa 3721 acaatctcat atgaagtcac cctagccatc attctactat caacattact aataagtggc 3781 tcctttaacc tctccaccct tatcacaaca caagaacacc tctgattact cctgccatca 3841 tgacccttgg ccataatatg atttatctcc acactagcag agaccaaccg aacccccttc 3901 gaccttgccg aaggggagtc cgaactagtc tcaggcttca acatcgaata cgccgcaggc 3961 cccttcgccc tattcttcat agccgaatac acaaacatta ttataataaa caccctcacc 4021 actacaatct tcctaggaac aacatatgac gcactctccc ctgaactcta cacaacatat 4081 tttgtcacca agaccctact tctaacctcc ctgttcttat gaattcgaac agcatacccc 4141 cgattccgct acgaccaact catacacctc ctatgaaaaa acttcctacc actcacccta 4201 gcattactta tatgatatgt ctccataccc attacaatct ccagcattcc ccctcaaacc 4261 taagaaatat gtctgataaa agagttactt tgatagagta aataatagga gcttaaaccc 4321 ccttatttct aggactatga gaatcgaacc catccctgag aatccaaaat tctccgtgcc 4381 acctatcaca ccccatccta aagtaaggtc agctaaataa gctatcgggc ccataccccg 4441 aaaatgttgg ttataccctt cccgtactaa ttaatcccct ggcccaaccc gtcatctact 4501 ctaccatctt tgcaggcaca ctcatcacag cgctaagctc gcactgattt tttacctgag 4561 taggcctaga aataaacatg ctagctttta ttccagttct aaccaaaaaa ataaaccctc 4621 gttccacaga agctgccatc aagtatttcc tcacgcaagc aaccgcatcc ataatccttc 4681 taatagctat cctcttcaac aatatactct ccggacaatg aaccataacc aatactacca 4741 atcaatactc atcattaata atcataatag ctatagcaat aaaactagga atagccccct 4801 ttcacttctg agtcccagag gttacccaag gcacccctct gacatccggc ctgcttcttc 4861 tcacatgaca aaaactagcc cccatctcaa tcatatacca aatctctccc tcactaaacg 4921 taagccttct cctcactctc tcaatcttat ccatcatagc aggcagttga ggtggattaa 4981 accaaaccca gctacgcaaa atcttagcat actcctcaat tacccacata ggatgaataa 5041 tagcagttct accgtacaac cctaacataa ccattcttaa tttaactatt tatattatcc 5101 taactactac cgcattccta ctactcaact taaactccag caccacgacc ctactactat 5161 ctcgcacctg aaacaagcta acatgactaa cacccttaat tccatccacc ctcctctccc 5221 taggaggcct gcccccgcta accggctttt tgcccaaatg ggccattatc gaagaattca 5281 caaaaaacaa tagcctcatc atccccacca tcatagccac catcaccctc cttaacctct 5341 acttctacct acgcctaatc tactccacct caatcacact actccccata tctaacaacg 5401 taaaaataaa atgacagttt gaacatacaa aacccacccc attcctcccc acactcatcg 5461 cccttaccac gctactccta cctatctccc cttttatact aataatctta tagaaattta 5521 ggttaaatac agaccaagag ccttcaaagc cctcagtaag ttgcaatact taatttctgt 5581 aacagctaag gactgcaaaa ccccactctg catcaactga acgcaaatca gccactttaa 5641 ttaagctaag cccttactag accaatggga cttaaaccca caaacactta gttaacagct 5701 aagcacccta atcaactggc ttcaatctac ttctcccgcc gccgggaaaa aaggcgggag 5761 aagccccggc aggtttgaag ctgcttcttc gaatttgcaa ttcaatatga aaatcacctc 5821 ggagctggta aaaagaggcc taacccctgt ctttagattt acagtccaat gcttcactca 5881 gccattttac ctcaccccca ctgatgttcg ccgaccgttg actattctct acaaaccaca 5941 aagacattgg aacactatac ctattattcg gcgcatgagc tggagtccta ggcacagctc 6001 taagcctcct tattcgagcc gagctgggcc agccaggcaa ccttctaggt aacgaccaca 6061 tctacaacgt tatcgtcaca gcccatgcat ttgtaataat cttcttcata gtaataccca 6121 tcataatcgg aggctttggc aactgactag ttcccctaat aatcggtgcc cccgatatgg 6181 cgtttccccg cataaacaac ataagcttct gactcttacc tccctctctc ctactcctgc 6241 tcgcatctgc tatagtggag gccggagcag gaacaggttg aacagtctac cctcccttag 6301 cagggaacta ctcccaccct ggagcctccg tagacctaac catcttctcc ttacacctag 6361 caggtgtctc ctctatctta ggggccatca atttcatcac aacaattatc aatataaaac 6421 cccctgccat aacccaatac caaacgcccc tcttcgtctg atccgtccta atcacagcag 6481 tcctacttct cctatctctc ccagtcctag ctgctggcat cactatacta ctaacagacc 6541 gcaacctcaa caccaccttc ttcgaccccg ccggaggagg agaccccatt ctataccaac 6601 acctattctg atttttcggt caccctgaag tttatattct tatcctacca ggcttcggaa 6661 taatctccca tattgtaact tactactccg gaaaaaaaga accatttgga tacataggta 6721 tggtctgagc tatgatatca attggcttcc tagggtttat cgtgtgagca caccatatat 6781 ttacagtagg aatagacgta gacacacgag catatttcac ctccgctacc ataatcatcg 6841 ctatccccac cggcgtcaaa gtatttagct gactcgccac actccacgga agcaatatga 6901 aatgatctgc tgcagtgctc tgagccctag gattcatctt tcttttcacc gtaggtggcc 6961 tgactggcat tgtattagca aactcatcac tagacatcgt actacacgac acgtactacg 7021 ttgtagccca cttccactat gtcctatcaa taggagctgt atttgccatc ataggaggct 7081 tcattcactg atttccccta ttctcaggct acaccctaga ccaaacctac gccaaaatcc 7141 atttcactat catattcatc ggcgtaaatc taactttctt cccacaacac tttctcggcc 7201 tatccggaat gccccgacgt tactcggact accccgatgc atacaccaca tgaaacatcc 7261 tatcatctgt aggctcattc atttctctaa cagcagtaat attaataatt ttcatgattt 7321 gagaagcctt cgcttcgaag cgaaaagtcc taatagtaga agaaccctcc ataaacctgg 7381 agtgactata tggatgcccc ccaccctacc acacattcga agaacccgta tacataaaat

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BIOL3251 GENETICS LAB FALL 2018

7441 ctagacaaaa aaggaaggaa tcgaaccccc caaagctggt ttcaagccaa ccccatggcc 7501 tccatgactt tttcaaaaag gtattagaaa aaccatttca taactttgtc aaagttaaat 7561 tataggctaa atcctatata tcttaatggc acatgcagcg caagtaggtc tacaagacgc 7621 tacttcccct atcatagaag agcttatcac ctttcatgat cacgccctca taatcatttt 7681 ccttatctgc ttcctagtcc tgtatgccct tttcctaaca ctcacaacaa aactaactaa 7741 tactaacatc tcagacgctc aggaaataga aaccgtctga actatcctgc ccgccatcat 7801 cctagtcctc atcgccctcc catccctacg catcctttac ataacagacg aggtcaacga 7861 tccctccctt accatcaaat caattggcca ccaatggtac tgaacctacg agtacaccga 7921 ctacggcgga ctaatcttca actcctacat acttccccca ttattcctag aaccaggcga 7981 cctgcgactc cttgacgttg acaatcgagt agtactcccg attgaagccc ccattcgtat 8041 aataattaca tcacaagacg tcttgcactc atgagctgtc cccacattag gcttaaaaac 8101 agatgcaatt cccggacgtc taaaccaaac cactttcacc gctacacgac cgggggtata 8161 ctacggtcaa tgctctgaaa tctgtggagc aaaccacagt ttcatgccca tcgtcctaga 8221 attaattccc ctaaaaatct ttgaaatagg gcccgtattt accctatagc accccctcta 8281 ccccctctag agcccactgt aaagctaact tagcattaac cttttaagtt aaagattaag 8341 agaaccaaca cctctttaca gtgaaatgcc ccaactaaat actaccgtat ggcccaccat 8401 aattaccccc atactcctta cactattcct catcacccaa ctaaaaatat taaacacaaa 8461 ctaccaccta cctccctcac caaagcccat aaaaataaaa aattataaca aaccctgaga 8521 accaaaatga acgaaaatct gttcgcttca ttcattgccc ccacaatcct aggcctaccc 8581 gccgcagtac tgatcattct atttccccct ctattgatcc ccacctccaa atatctcatc 8641 aacaaccgac taatcaccac ccaacaatga ctaatcaaac taacctcaaa acaaatgata 8701 accatacaca acactaaagg acgaacctga tctcttatac tagtatcctt aatcattttt 8761 attgccacaa ctaacctcct cggactcctg cctcactcat ttacaccaac cacccaacta 8821 tctataaacc tagccatggc catcccctta tgagcgggca cagtgattat aggctttcgc 8881 tctaagatta aaaatgccct agcccacttc ttaccacaag gcacacctac accccttatc 8941 cccatactag ttattatcga aaccatcagc ctactcattc aaccaatagc cctggccgta 9001 cgcctaaccg ctaacattac tgcaggccac ctactcatgc acctaattgg aagcgccacc 9061 ctagcaatat caaccattaa ccttccctct acacttatca tcttcacaat tctaattcta 9121 ctgactatcc tagaaatcgc tgtcgcctta atccaagcct acgttttcac acttctagta 9181 agcctctacc tgcacgacaa cacataatga cccaccaatc acatgcctat catatagtaa 9241 aacccagccc atgaccccta acaggggccc tctcagccct cctaatgacc tccggcctag 9301 ccatgtgatt tcacttccac tccataacgc tcctcatact aggcctacta accaacacac 9361 taaccatata ccaatgatgg cgcgatgtaa cacgagaaag cacataccaa ggccaccaca 9421 caccacctgt ccaaaaaggc cttcgatacg ggataatcct atttattacc tcagaagttt 9481 ttttcttcgc aggatttttc tgagcctttt accactccag cctagcccct accccccaat 9541 taggagggca ctggccccca acaggcatca ccccgctaaa tcccctagaa gtcccactcc 9601 taaacacatc cgtattactc gcatcaggag tatcaatcac ctgagctcac catagtctaa 9661 tagaaaacaa ccgaaaccaa ataattcaag cactgcttat tacaatttta ctgggtctct 9721 attttaccct cctacaagcc tcagagtact tcgagtctcc cttcaccatt tccgacggca 9781 tctacggctc aacatttttt gtagccacag gcttccacgg acttcacgtc attattggct 9841 caactttcct cactatctgc ttcatccgcc aactaatatt tcactttaca tccaaacatc 9901 actttggctt cgaagccgcc gcctgatact ggcattttgt agatgtggtt tgactatttc 9961 tgtatgtctc catctattga tgagggtctt actcttttag tataaatagt accgttaact 10021 tccaattaac tagttttgac aacattcaaa aaagagtaat aaacttcgcc ttaattttaa 10081 taatcaacac cctcctagcc ttactactaa taattattac attttgacta ccacaactca 10141 acggctacat agaaaaatcc accccttacg agtgcggctt cgaccctata tcccccgccc 10201 gcgtcccttt ctccataaaa ttcttcttag tagctattac cttcttatta tttgatctag 10261 aaattgccct ccttttaccc ctaccatgag ccctacaaac aactaacctg ccactaatag 10321 ttatgtcatc cctcttatta atcatcatcc tagccctaag tctggcctat gagtgactac 10381 aaaaaggatt agactgaacc gaattggtat atagtttaaa caaaacgaat gatttcgact 10441 cattaaatta tgataatcat atttaccaaa tgcccctcat ttacataaat attatactag 10501 catttaccat ctcacttcta ggaatactag tatatcgctc acacctcata tcctccctac 10561 tatgcctaga aggaataata ctatcgctgt tcattatagc tactctcata accctcaaca 10621 cccactccct cttagccaat attgtgccta ttgccatact agtctttgcc gcctgcgaag 10681 cagcggtggg cctagcccta ctagtctcaa tctccaacac atatggccta gactacgtac 10741 ataacctaaa cctactccaa tgctaaaact aatcgtccca acaattatat tactaccact 10801 gacatgactt tccaaaaaac acataatttg aatcaacaca accacccaca gcctaattat 10861 tagcatcatc cctctactat tttttaacca aatcaacaac aacctattta gctgttcccc 10921 aaccttttcc tccgaccccc taacaacccc cctcctaata ctaactacct gactcctacc 10981 cctcacaatc atggcaagcc aacgccactt atccagtgaa ccactatcac gaaaaaaact 11041 ctacctctct atactaatct ccctacaaat ctccttaatt ataacattca cagccacaga 11101 actaatcata ttttatatct tcttcgaaac cacacttatc cccaccttgg ctatcatcac 11161 ccgatgaggc aaccagccag aacgcctgaa cgcaggcaca tacttcctat tctacaccct 11221 agtaggctcc cttcccctac tcatcgcact aatttacact cacaacaccc taggctcact

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BIOL3251 GENETICS LAB FALL 2018

11281 aaacattcta ctactcactc tcactgccca agaactatca aactcctgag ccaacaactt 11341 aatatgacta gcttacacaa tagcttttat agtaaagata cctctttacg gactccactt 11401 atgactccct aaagcccatg tcgaagcccc catcgctggg tcaatagtac ttgccgcagt 11461 actcttaaaa ctaggcggct atggtataat acgcctcaca ctcattctca accccctgac 11521 aaaacacata gcctacccct tccttgtact atccctatga ggcataatta taacaagctc 11581 catctgccta cgacaaacag acctaaaatc gctcattgca tactcttcaa tcagccacat 11641 agccctcgta gtaacagcca ttctcatcca aaccccctga agcttcaccg gcgcagtcat 11701 tctcataatc gcccacgggc ttacatcctc attactattc tgcctagcaa actcaaacta 11761 cgaacgcact cacagtcgca tcataatcct ctctcaagga cttcaaactc tactcccact 11821 aatagctttt tgatgacttc tagcaagcct cgctaacctc gccttacccc ccactattaa 11881 cctactggga gaactctctg tgctagtaac cacgttctcc tgatcaaata tcactctcct 11941 acttacagga ctcaacatac tagtcacagc cctatactcc ctctacatat ttaccacaac 12001 acaatggggc tcactcaccc accacattaa caacataaaa ccctcattca cacgagaaaa 12061 caccctcatg ttcatacacc tatcccccat tctcctccta tccctcaacc ccgacatcat 12121 taccgggttt tcctcttgta aatatagttt aaccaaaaca tcagattgtg aatctgacaa 12181 cagaggctta cgacccctta tttaccgaga aagctcacaa gaactgctaa ctcatgcccc 12241 catgtctaac aacatggctt tctcaacttt taaaggataa cagctatcca ttggtcttag 12301 gccccaaaaa ttttggtgca actccaaata aaagtaataa ccatgcacac tactataacc 12361 accctaaccc tgacttccct aattcccccc atccttacca ccctcgttaa ccctaacaaa 12421 aaaaactcat acccccatta tgtaaaatcc attgtcgcat ccacctttat tatcagtctc 12481 ttccccacaa caatattcat gtgcctagac caagaagtta ttatctcgaa ctgacactga 12541 gccacaaccc aaacaaccca gctctcccta agcttcaaac tagactactt ctccataata 12601 ttcatccctg tagcattgtt cgttacatgg tccatcatag aattctcact gtgatatata 12661 aactcagacc caaacattaa tcagttcttc aaatatctac tcatcttcct aattaccata 12721 ctaatcttag ttaccgctaa caacctattc caactgttca tcggctgaga gggcgtagga 12781 attatatcct tcttgctcat cagttgatga tacgcccgag cagatgccaa cacagcagcc 12841 attcaagcaa tcctatacaa ccgtatcggc gatatcggtt tcatcctcgc cttagcatga 12901 tttatcctac actccaactc atgagaccca caacaaatag cccttctaaa cgctaatcca 12961 agcctcaccc cactactagg cctcctccta gcagcagcag gcaaatcagc ccaattaggt 13021 ctccacccct gactcccctc agccatagaa ggccccaccc cagtctcagc cctactccac 13081 tcaagcacta tagttgtagc aggaatcttc ttactcatcc gcttccaccc cctagcagaa 13141 aatagcccac taatccaaac tctaacacta tgcttaggcg ctatcaccac tctgttcgca 13201 gcagtctgcg cccttacaca aaatgacatc aaaaaaatcg tagccttctc cacttcaagt 13261 caactaggac tcataatagt tacaatcggc atcaaccaac cacacctagc attcctgcac 13321 atctgtaccc acgccttctt caaagccata ctatttatgt gctccgggtc catcatccac 13381 aaccttaaca atgaacaaga tattcgaaaa ataggaggac tactcaaaac catacctctc 13441 acttcaacct ccctcaccat tggcagccta gcattagcag gaataccttt cctcacaggt 13501 ttctactcca aagaccacat catcgaaacc gcaaacatat catacacaaa cgcctgagcc 13561 ctatctatta ctctcatcgc tacctccctg acaagcgcct atagcactcg aataattctt 13621 ctcaccctaa caggtcaacc tcgcttcccc acccttacta acattaacga aaataacccc 13681 accctactaa accccattaa acgcctggca gccggaagcc tattcgcagg atttctcatt 13741 actaacaaca tttcccccgc atcccccttc caaacaacaa tccccctcta cctaaaactc 13801 acagccctcg ctgtcacttt cctaggactt ctaacagccc tagacctcaa ctacctaacc 13861 aacaaactta aaataaaatc cccactatgc acattttatt tctccaacat actcggattc 13921 taccctagca tcacacaccg cacaatcccc tatctaggcc ttcttacgag ccaaaacctg 13981 cccctactcc tcctagacct aacctgacta gaaaagctat tacctaaaac aatttcacag 14041 caccaaatct ccacctccat catcacctca acccaaaaag gcataattaa actttacttc 14101 ctctctttct tcttcccact catcctaacc ctactcctaa tcacataacc tattcccccg 14161 agcaatctca attacaatat atacaccaac aaacaatgtt caaccagtaa ctactactaa 14221 tcaacgccca taatcataca aagcccccgc accaatagga tcctcccgaa tcaaccctga 14281 cccctctcct tcataaatta ttcagcttcc tacactatta aagtttacca caaccaccac 14341 cccatcatac tctttcaccc acagcaccaa tcctacctcc atcgctaacc ccactaaaac 14401 actcaccaag acctcaaccc ctgaccccca tgcctcagga tactcctcaa tagccatcgc 14461 tgtagtatat ccaaagacaa ccatcattcc ccctaaataa attaaaaaaa ctattaaacc 14521 catataacct cccccaaaat tcagaataat aacacacccg accacaccgc taacaatcaa 14581 tactaaaccc ccataaatag gagaaggctt agaagaaaac cccacaaacc ccattactaa 14641 acccacactc aacagaaaca aagcatacat cattattctc gcacggacta caaccacgac 14701 caatgatatg aaaaaccatc gttgtatttc aactacaaga acaccaatga ccccaatacg 14761 caaaactaac cccctaataa aattaattaa ccactcattc atcgacctcc ccaccccatc 14821 caacatctcc gcatgatgaa acttcggctc actccttggc gcctgcctga tcctccaaat 14881 caccacagga ctattcctag ccatgcacta ctcaccagac gcctcaaccg ccttttcatc 14941 aatcgcccac atcactcgag acgtaaatta tggctgaatc atccgctacc ttcacgccaa 15001 tggcgcctca atattcttta tctgcctctt cctacacatc gggcgaggcc tatattacgg 15061 atcatttctc tactcagaaa cctgaaacat cggcattatc ctcctgcttg caactatagc

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BIOL3251 GENETICS LAB FALL 2018

15121 aacagccttc ataggctatg tcctcccgtg aggccaaata tcattctgag gggccacagt 15181 aattacaaac ttactatccg ccatcccata cattgggaca gacctagttc aatgaatctg 15241 aggaggctac tcagtagaca gtcccaccct cacacgattc tttacctttc acttcatctt 15301 gcccttcatt attgcagccc tagcaacact ccacctccta ttcttgcacg aaacgggatc 15361 aaacaacccc ctaggaatca cctcccattc cgataaaatc accttccacc cttactacac 15421 aatcaaagac gccctcggct tacttctctt ccttctctcc ttaatgacat taacactatt 15481 ctcaccagac ctcctaggcg acccagacaa ttatacccta gccaacccct taaacacccc 15541 tccccacatc aagcccgaat gatatttcct attcgcctac acaattctcc gatccgtccc 15601 taacaaacta ggaggcgtcc ttgccctatt actatccatc ctcatcctag caataatccc 15661 catcctccat atatccaaac aacaaagcat aatatttcgc ccactaagcc aatcacttta 15721 ttgactccta gccgcagacc tcctcattct aacctgaatc ggaggacaac cagtaagcta 15781 cccttttacc atcattggac aagtagcatc cgtactatac ttcacaacaa tcctaatcct 15841 aataccaact atctccctaa ttgaaaacaa aatactcaaa tgggcctgtc cttgtagtat 15901 aaactaatac accagtcttg taaaccggag atgaaaacct ttttccaagg acaaatcaga 15961 gaaaaagtct ttaactccac cattagcacc caaagctaag attctaattt aaactattct 16021 ctgttctttc atggggaagc agatttgggt accacccaag tattgactca cccatcaaca 16081 accgctatgt atttcgtaca ttactgccag ccaccatgaa tattgtacgg taccataaat 16141 acttgaccac ctgtagtaca taaaaaccca atccacatca aaaccccctc cccatgctta 16201 caagcaagta cagcaatcaa ccctcaacta tcacacatca actgcaactc caaagccacc 16261 cctcacccac taggatacca acaaacctac ccacccttaa cagtacatag tacataaagc 16321 catttaccgt acatagcaca ttacagtcaa atcccttctc gtccccatgg atgacccccc 16381 tcagataggg gtcccttgac caccatcctc cgtgaaatca atatcccgca caagagtgct 16441 actctcctcg ctccgggccc ataacacttg ggggtagcta aagtgaactg tatccgacat 16501 ctggttccta cttcagggtc ataaagccta aatagcccac acgttcccct taaataagac 16561 atcacgatg

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